Inhibitors of short-chain dehydrogenase activity for modulating hematopoietic stem cells and hematopoiesis

ABSTRACT

A method of modulating hematopoietic stem cells and hematopoiesis includes administering to a subject in need thereof a 15-PGDH inhibitor.

RELATED APPLICATION

This application is a Continuation-in-part of U.S. patent applicationSer. No. 14/395,021, filed Oct. 16, 2014, which is a National PhaseFiling of PCT/US2013/036790, filed Apr. 16, 2013, which claims priorityto U.S. Provisional Application Ser. No. 61/624,670, filed Apr. 16,2012, this application is also a Continuation-in-Part of U.S. Ser. No.15/029,943, filed Apr. 15, 2016, which is a National Phase Filing ofPCT/US2014/060761, filed Oct. 16, 2014, which claims priority to U.S.Provisional Application Nos. 61/891,260, filed Oct. 15, 2013,61/954,202, filed Mar. 17, 2014, 62/019,597, filed Jul. 1, 2014, and62/043,694, filed Aug. 29, 2014, this application also claims priorityto PCT/US2016/027549, filed Apr. 14, 2016, which claims priority to U.S.Provisional Application No. 62/129,885, filed Mar. 8, 2015, and thisapplication also claims priority from U.S. Provisional Application No.62/252,973, filed Nov. 9, 2015, the subject matter of which isincorporated herein by reference in its entirety.

GOVERNMENT FUNDING

This invention was made with government support under Grant No.R01CA127306, R01CA127306-0351, 1P01CA95471-10, AND 5P50CA150964 awardedby The National Institutes of Health. The United States government hascertain rights in the invention.

BACKGROUND

Short-chain dehydrogenases (SCDs) are a family of dehydrogenases thatshare only 15% to 30% sequence identity, with similarity predominantlyin the coenzyme binding domain and the substrate binding domain. Inaddition to their role in detoxification of ethanol, SCDs are involvedin synthesis and degradation of fatty acids, steroids, and someprostaglandins, and are therefore implicated in a variety of disorders,such as lipid storage disease, myopathy, SCD deficiency, and certaingenetic disorders.

The SCD, 15-hydroxy-prostaglandin dehydrogenase (15-PGDH),(hydroxyprostaglandin dehydrogenase 15-(nicotinamideadeninedinucleotide); 15-PGDH; Enzyme Commission number 1.1.1.141;encoded by the HPGD gene), represents the key enzyme in the inactivationof a number of active prostaglandins, leukotrienes andhydroxyeicosatetraenoic acids (HETEs) (e.g., by catalyzing oxidation ofPGE₂ to 15-keto-prostaglandin E2, 15k-PGE). The human enzyme is encodedby the HPGD gene and consists of a homodimer with subunits of a size of29 kDa. The enzyme belongs to the evolutionarily conserved superfamilyof short-chain dehydrogenase/reductase enzymes (SDRs), and according tothe recently approved nomenclature for human enzymes, it is namedSDR36C1. Thus far, two forms of 15-PGDH enzyme activity have beenidentified, NAD+-dependent type I 15-PGDH that is encoded by the HPGDgene, and the type II NADP-dependent 15-PGDH, also known as carbonylreductase 1 (CBR1, SDR21C1). However, the preference of CBR1 for NADPand the high Km values of CBR1 for most prostaglandin suggest that themajority of the in vivo activity can be attributed to type I 15-PGDHencoded by the HPGD gene, that hereafter, and throughout all followingtext, simply denoted as 15-PGDH.

SUMMARY

Embodiments described herein relate to compositions and methods formodulating hematopoietic stem cells and hematopoiesis. As described inthe Examples below, it was found that that inhibitors of short-chaindehydrogenase activity, such as 15-PGDH inhibitors, can be administeredto a subject in need thereof, cells of the subject, and/or tissue of thesubject alone or in combination with a cytokine to increase and/ormobilize hematopoietic stem cells and/or neutrophils in the blood,marrow, and/or tissue of the subject.

In some embodiments, the administration of a 15-PGDH inhibitor alone orin combination with a cytokine, hematopoietic cytokine, and/or cellmobilization agent to a subject in need thereof, cells of the subject,and/or tissue of the subject can be used for the purpose of increasingneutrophils in the subject. For example, the hematopoietic cytokine orcell mobilization agent can include at least one of G-CSF or Plerixafor.

In other embodiments, the administration of a 15-PGDH inhibitor alone orin combination with a cytokine, hematopoietic cytokine, and/or cellmobilization agent to a subject in need thereof, cells of the subject,and/or tissue of the subject can be used for the purpose of increasingnumbers of and/or of mobilizing peripheral blood hematopoietic stemcells in the subject. For example, the hematopoietic cytokine or cellmobilization agent can include at least one of G-CSF or Plerixafor.

In still other embodiments, the administration of a 15-PGDH inhibitoralone or in combination with a cytokine, hematopoietic cytokine, and/orcell mobilization agent to a subject in need thereof, cells of thesubject, and/or tissue of the subject can be used for the purpose ofincreasing numbers of hematopoietic stem cells in blood or bone marrowof the subject. For example, the hematopoietic cytokine or cellmobilization agent can include at least one of G-CSF or Plerixafor.

In other embodiments, the 15-PGDH inhibitor can be administered to asubject and/or tissue of the subject to increase tissue stem cells. Forexample, the 15-PGDH inhibitor can be administered to bone marrow of asubject to increase stem cells in the subject.

In still other embodiments, the 15-PGDH inhibitor can be administered toa tissue graft donor, bone marrow graft donor, and/or a hematopoieticstem cell donor, and/or a tissue graft, and/or a bone marrow graft,and/or a hematopoietic stem cell graft, to increase the fitness of adonor tissue graft, a donor bone marrow graft, and/or a donorhematopoietic stem cell graft. For example, the 15-PGDH inhibitor can beadministered to a subject, and/or bone marrow of a subject to increasethe fitness of the marrow as a donor graft, and/or to a preparation ofhematopoietic stem cells of a subject to increase the fitness of thestem cell preparation as a donor graft, and/or to a preparation ofperipheral blood hematopoietic stem cells of a subject to increase thefitness of the stem cell preparation as a donor graft, and/or to apreparation of umbilical cord blood stem cells to increase the fitnessof the stem cell preparation as a donor graft, and/or to a preparationof umbilical cord blood stem cells to decrease the number of units ofumbilical cord blood required for transplantation.

In other embodiments, the 15-PGDH inhibitor can be administered to arecipient of a tissue graft transplant, bone marrow transplant,hematopoietic stem cell transplant, and/or of an umbilical cord stemcell transplant, in order to decrease the administration of othertreatments or growth factors.

In still other embodiments, the 15-PGDH inhibitor can be administered toa subject to increase neutrophil counts following a hematopoietic celltransplant with bone marrow, hematopoietic stem cells, or umbilical cordblood, to increase neutrophil counts in a subject with neutropeniafollowing chemotherapy administration or radiation therapy, to increaseneutrophil counts in a subject with aplastic anemia, myelodysplasia,myelofibrosis, neutropenia due to other bone marrow diseases, druginduced neutropenia, autoimmune neutropenia, idiopathic neutropenia, orneutropenia following viral infections, to increase neutrophil counts ina subject with neutropenia, to increase platelet counts following ahematopoietic cell transplant with bone marrow, hematopoietic stemcells, or umbilical cord blood, to increase platelet counts in a subjectwith thrombocytopenia following chemotherapy administration or radiationtherapy, to increase platelet counts in a subject with aplastic anemia,myelodysplasia, myelofibrosis, thrombocytopenia due to other bone marrowdiseases, drug induced thrombocytopenia, autoimmune thrombocytopenia,idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia, orthrombocytopenia following viral infections, to increase platelet countsin a subject with thrombocytopenia, to increase red blood cell counts,or hematocrit, or hemoglobin level, following a hematopoietic celltransplant with bone marrow, hematopoietic stem cells, or umbilical cordblood, to increase red blood cell counts, or hematocrit, or hemoglobinlevel in a subject with anemia following chemotherapy administration orradiation therapy, to increase red blood cell counts, or hematocrit, orhemoglobin level counts in a subject with aplastic anemia,myelodysplasia, myelofibrosis, anemia due to other disorder of bonemarrow, drug induced anemia, immune mediated anemias, anemia of chronicdisease, anemia following viral infections, or anemia of unknown cause,to increase red blood cell counts, or hematocrit, or hemoglobin level ina subject with anemia, to increase bone marrow stem cells, following ahematopoietic cell transplant with bone marrow, hematopoietic stemcells, or umbilical cord blood, to increase bone marrow stem cells in asubject following chemotherapy administration or radiation therapy,and/or to increase bone marrow stem cells in a subject with aplasticanemia, myelodysplasia, myelofibrosis, other disorder of bone marrow,drug induced cytopenias, immune cytopenias, cytopenias following viralinfections, or cytopenias.

In other embodiments, the 15-PGDH inhibitor can be administered to asubject having or at risk of presence of cytopenia, neutropenia,thrombocytopenia, lymphocytopenia and anemia to increase responsivenessand/or potentiate cytokines, such as hematopoietic cytokines. Thecytokines can include, for example, G-CSF, GM-CSF, EPO, IL-3, IL-6, TPO,TPO-RA (thrombopoietin receptor agonist), and SCF.

In some embodiments, the subject has received a hematopoietic stem celltransplant, bone marrow transplant, chemotherapy, a myelosuppressivetherapy, radiation therapy, or viral therapy and/or has a bone marrowdisease, cancer, viral infection, aplastic anemia, myelodysplasia,and/or myelofibrosis.

Other embodiments relate to a method of treating at least one ofneutropenia, thrombocytopenia, anemia, or cytopenia in a subject in needthereof by administering to the subject a therapeutically effectiveamount of a15-PGDH inhibitor. The 15-PGDH inhibitor can be administeredalone or in combination with a cytokine, hematopoietic cytokine, and/orcell mobilization agent to the subject, cells of the subject, and/ortissue of the subject.

In some embodiments, the 15-PGDH inhibitor can be administered to ablood, bone marrow, and/or tissue of a subject at an amount effective toincrease prostaglandin levels in the subject. The 15-PGDH inhibitor caninclude a compound having formula (I):

wherein n is 0-2;

Y¹, Y², and R¹ are the same or different and are each selected from thegroup consisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O) (C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), phosphono (—P(O)(OH)₂), phosphonato(—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂), phosphino (—PH₂),combinations thereof, and wherein Y¹ and Y² may be linked to form acyclic or polycyclic ring, wherein the ring is a substituted orunsubstituted aryl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted cycloalkyl, and a substituted orunsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ aresame or different and are each selected from the group consisting of H,a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

X¹ and X² are independently N or C, and wherein when X¹ and/or X² are N,Y¹ and/or Y², respectively, are absent;

Z¹ is O, S, CR^(a)R^(b) or NR^(a), wherein R^(a) and R^(b) areindependently H or a C₁₋₈ alkyl, which is linear, branched, or cyclic,and which is unsubstituted or substituted;

and pharmaceutically acceptable salts thereof.

In other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (V):

wherein n is 0-2

X⁶ is independently is N or CR^(c)

R¹, R⁶, R⁷, and R^(c) are each independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O)(C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinR⁶ and R⁷ may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ arethe same or different and are each selected from the group consisting ofH, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

and pharmaceutically acceptable salts thereof.

In some embodiments, R¹ is selected from the group consisting ofbranched or linear alkyl including —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5).

In other embodiments, R⁶ and R⁷ can each independently be one of thefollowing:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, and R⁷⁴, are the sameor different and are independently selected from the group consisting ofhydrogen, substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heterocycloalkenyl containing from 5-6 ringatoms, (wherein from 1-3 of the ring atoms is independently selectedfrom N, NH, N(C₁-C₆ alkyl), NC(O)(C₁-C₆ alkyl), O, and S), heteroaryl orheterocyclyl containing from 5-14 ring atoms, (wherein from 1-6 of thering atoms is independently selected from N, NH, N(C₁-C₃ alkyl), O, andS), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, silyl, hydroxyl, sulfhydryl,C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy,acyl (including C₂-C₂₄ alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl(—CO-aryl)), acyloxy (—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl),C₆-C₂₀ aryloxycarbonyl (—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato(—O—(CO)—O-alkyl), C₆-C₂₀ arylcarbonato (—O—(CO)—O-aryl), carboxy(—COOH), carboxylato (—COO⁻), carbamoyl (—(CO)—NH₂), C₁-C₂₄alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)), arylcarbamoyl (—(CO)—NH-aryl),thiocarbamoyl (—(CS)—NH₂), carbamido (—NH—(CO)—NH₂), cyano(-CN),isocyano (—N⁺C⁻), cyanato (—O—CN), isocyanato (—O—N⁺═C⁻), isothiocyanato(—S—CN), azido (—N═N⁺═N⁻), formyl (—(CO)—H), thioformyl (—(CS)—H), amino(—NH₂), C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido(—NH—(CO)-alkyl), C₆-C₂₀ arylamido (—NH—(CO)-aryl), sulfanamido(—SO₂N(R)₂ where R is independently H, alkyl, aryl or heteroaryl), imino(—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen,alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), whereR=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO₂), nitroso (—NO),sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄ alkylsulfanyl (—S-alkyl;also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed“arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl), C₅-C₂₀ arylsulfinyl(—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl), C₅-C₂₀ arylsulfonyl(—SO₂-aryl), sulfonamide (—SO₂—NH₂, —SO₂NY₂ (wherein Y is independentlyH, aryl or alkyl), phosphono (—P(O)(OH)₂), phosphonato (—P(O)(O⁻)₂),phosphinato (—P(O)(O⁻)), phospho (—PO₂), phosphino (—PH₂), polyalkylethers (—[(CH₂)_(n)O]_(m)), phosphates, phosphate esters [—OP(O)(OR)₂where R═H, methyl or other alkyl], groups incorporating amino acids orother moieties expected to bear positive or negative charge atphysiological pH, and combinations thereof, and pharmaceuticallyacceptable salts thereof.

In some embodiments, the 15-PGDH inhibitor can inhibit the enzymaticactivity of recombinant 15-PGDH at an IC₅₀ of less than 1 μM, orpreferably at an IC₅₀ of less than 250 nM, or more preferably at an IC₅₀of less than 50 nM, or more preferably at an IC₅₀ of less than 10 nM, ormore preferably at an IC₅₀ of less than 5 nM at a recombinant 15-PGDHconcentration of about 5 nM to about 10 nM.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a graph showing CFU counts in wild type bone marrowtreated with SW033291 and PGE-2.

FIGS. 2(A-F) illustrate graphs showing: (A) bone marrow cellularity ofmice treated with SW033291; (B) white blood cell (WBC) counts in micetreated SW033291; (C) red blood cell (RBC) counts in mice treatedSW033291; (D) platelet counts in mice treated SW033291; (E) SKL % inwhole bone marrow of mice treated with SW033291; and (F) CFU counts inmice treated SW033291.

FIGS. 3(A-B) illustrate: (A) a schematic diagram following CD45.2antigen marked cells in lethally irradiated C57BL/6J mice rescued with abone marrow transplant from donor mice treated with SW033291 or withvehicle; and (B) graphs showing chimerism, of donor B-Cells, myeloidcells, and T-Cells after such treatment.

FIG. 4 illustrates a schematic diagram showing schema of a study inwhich C57BL/61 mice are irradiated with 11GY on day 0 and followed bytreatment with SW033291.

FIGS. 5(A-D) illustrate: (A) a schematic illustration showing the designof a study of enhanced survival in mice receiving a bone marrowtransplant and also administered the 15-PGDH inhibitor SW033291; (B)graphical survival curves for mice transplanted with 100,000 donorcells; (C) graphical survival curves for mice transplanted with 200,000donor cells; and (D) graphical survival curves for mice transplantedwith 500,000 donor cells.

FIGS. 6(A-C) illustrate: (A) a schematic illustration showingmeasurements on blood and bone marrow on day 5 after transplant; (B) agraph showing that SW033291 treated mice have significantly higher totalwhite count; (C) a graph showing that SW033291 treated mice havesignificantly higher total platelet count. The star symbol denotesP<0.05.

FIGS. 7(A-B) illustrate: (A) a schematic illustration showingmeasurements on blood and bone marrow on day 8 after transplant; and (B)a graph showing that SW033291 treated mice have significantly higherplatelet count than control, with drug treated mice having 77,000platelets compared to control mice having 39,500 platelets. The starsymbol denotes P<0.05.

FIGS. 8(A-D) illustrate: (A) a schematic illustration showingmeasurements on blood and bone marrow on day 12 after transplant; (B) agraph showing that SW033291 treated mice have significantly higherneutrophil counts, with drug treated mice having 332 neutrophilscompared to control mice having 125 neutrophils; and (C) a graph showingthat on day 12 after transplant, SW033291 treated mice havesignificantly higher hemoglobin count than controls, with drug treatedmice having hemoglobin level of 11.58 and control mice having hemoglobinlevel of 8.3; and D) a graph showing that SW033291 treated mice havesignificantly higher total white counts compared to control mice. Thestar symbol denotes P<0.05.

FIGS. 9(A-G) illustrate: (A) a schematic illustration showingmeasurements on blood and bone marrow on day 18 after transplant; (B-D)graphs showing SW033291 treated mice have significantly higher totalwhite count (FIG. 85B), lymphocyte count (FIG. 85C), and neutrophilcount (FIG. 85D), with drug treated mice having 835 neutrophils andcontrol mice having 365 neutrophils (FIG. 85D); (E) a graph showing thaton day 18 drug treated mice have significantly higher platelet countsthan control mice; and (F-G) graphs showing drug treated mice havenearly 4-fold increased percentage (FIG. 85F) and total numbers (FIG.85G) of SKL marked bone marrow stem cells than do control mice. The starsymbol denotes P<0.05.

FIGS. 10(A-B) illustrate graphs showing (A) measurement of PGE2 (pg ofPGE2/mg tissue protein) in 4 different mouse tissues (colon, bonemarrow, liver, lung) across time following IP injection of SW033291 at10 mg/kg; and (B) time course of PGE2 in control mice injected withvehicle only.

FIG. 11 is a schematic illustration showing an experiment in which miceare lethally irradiated (IR) and 12 hours later receive a transplant(BMT) with CFSE dye labeled bone marrow cells (BM), and the number oftransplanted cells that home and survive in the bone marrow of therecipient mice are then determined by FACS at 16 hours post-transplant.

FIG. 12 illustrates a graph showing the percent of CFSE dye labeledcells that have homed to the bone marrow of mice treated as illustratedin FIG. 11.

FIG. 13 is a schematic illustration showing an experiment in which miceare lethally irradiated (IR) and 12 hours later receive a transplant(BMT) with CFSE dye labeled bone marrow cells (BM), and numbers oftransplanted cells that home and survive in the bone marrow of therecipient mice are then determined by FACS at 16 hours post-transplant.

FIG. 14 illustrates a graph showing the percent of CFSE dye labeledcells that have homed to the bone marrow of mice treated as illustratedin FIG. 13.

FIGS. 15(A-B) illustrate graphs showing induction of gene expression in(A) bone marrow SKL cells and (B) bone marrow stromal cells of miceinjected with SW033291 twice daily IP at 10 mg/kg for 3 days.

FIG. 16 is a schematic illustration showing an experiment in whichimmune deficient NSG mice are lethally irradiated (IR) and 12 hourslater receive a transplant with CFSE dye labeled buffy coat cells fromhuman umbilical cord blood (UCB), and number of transplanted cells thathome and survive in the bone marrow of the recipient mice are thendetermined by FACS at 16 hours post-transplant.

FIG. 17 illustrates a graph showing the percent of CFSE dye labeledhuman umbilical cord buffy coat cells that have homed to the bone marrowof mice treated as per the schema above.

FIG. 18 illustrates a schematic diagram showing the design of a study inwhich 4 groups of mice (4 mice per group) were administered either A)vehicle control; B) 2.5 mg/kg of 15-PGDH inhibitor (+) SW209415 twicedaily IP for 7 doses; c) recombinant human G-CSF 250 μg/kgsubcutaneously once daily for 4 doses or d) the combination of 2.5 mg/kgof 15-PGDH inhibitor (+) SW209415 twice daily IP for 7 doses plusrecombinant human G-CSF 250 μg/kg subcutaneously once daily for 4 doses(with the daily dose of G-CSF being administered coincident with a doseof (+) SW209415).

FIG. 19 illustrates graphs showing total counts of white blood cells,neutrophils, and lymphocytes of mice administered 15-PGDH inhibitor (+)SW209415 and/or G-CSF as described in FIG. 18.

FIG. 20 illustrates graphs showing total counts of red blood cells,hematocrit, hemoglobin, and platelets of mice administered 15-PGDHinhibitor (+) SW209415 and/or G-CSF as described in FIG. 18.

FIG. 21 illustrates graphs showing the percent and number of circulatingSKL (Sca-1+; C-kit+; Lin−) marked cells in peripheral blood of miceadministered 15-PGDH inhibitor (+) SW209415 and/or G-CSF as described inFIG. 18.

FIG. 22 illustrates graphs showing the percent and number of SKL(Sca-1+; C-kit+; Lin−) marked cells in bone marrow of mice administered15-PGDH inhibitor (+) SW209415 and/or G-CSF as described in FIG. 18.

FIGS. 23(A-G) illustrate graphs showing (A) WBC counts, (B) neutrophilcounts, (C) lymphocyte counts, (D) RBC counts, (E) hemoglobin counts,(F) hematocrit counts, and (G) platelet counts in peripheral blood ofrecipient mice lethally irradiated followed by bone marrowtransplantation (BMT) treated with vehicle, G-CSF, SW209415, andSW209415 in combination with G-CSF at day 8 following BMT.

FIGS. 24(A-G) illustrate graphs showing (A) WBC counts, (B) neutrophilcounts, (C) lymphocyte counts, (D) RBC counts, (E) hemoglobin counts,(F) hematocrit counts, and (G) platelet counts in peripheral blood ofrecipient mice lethally irradiated followed by bone marrowtransplantation (BMT) treated with vehicle, G-CSF, SW209415, andSW209415 in combination with G-CSF at day 12 following BMT.

FIGS. 25(A-G) illustrate graphs showing (A) WBC counts, (B) neutrophilcounts, (C) lymphocyte counts, (D) RBC counts, (E) hemoglobin counts,(F) hematocrit counts, and (G) platelet counts in peripheral blood ofrecipient mice lethally irradiated followed by bone marrowtransplantation (BMT) treated with vehicle, G-CSF, SW209415, andSW209415 in combination with G-CSF at day 18 following BMT.

FIGS. 26(A-C) illustrate graphs showing (A) BM cellularity 18 days post500 k cell does BMY, (B) SKL levels 18 days post 500 k cell dose BMT,and (C) SKL/mouse 18 days post 500 k cell dose BMT treated with vehicle,G-CSF, SW209415, and SW209415 in combination with G-CSF at day 18following BMT.

DETAILED DESCRIPTION

For convenience, certain terms employed in the specification, examples,and appended claims are collected here. Unless defined otherwise, alltechnical and scientific terms used herein have the same meaning ascommonly understood by one of ordinary skill in the art to which thisapplication belongs.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

The terms “comprise,” “comprising,” “include,” “including,” “have,” and“having” are used in the inclusive, open sense, meaning that additionalelements may be included. The terms “such as”, “e.g.”, as used hereinare non-limiting and are for illustrative purposes only. “Including” and“including but not limited to” are used interchangeably.

The term “or” as used herein should be understood to mean “and/or”,unless the context clearly indicates otherwise.

As used herein, the term “about” or “approximately” refers to aquantity, level, value, number, frequency, percentage, dimension, size,amount, weight or length that varies by as much as 15%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2% or 1% to a reference quantity, level, value, number,frequency, percentage, dimension, size, amount, weight or length. In oneembodiment, the term “about” or “approximately” refers a range ofquantity, level, value, number, frequency, percentage, dimension, size,amount, weight or length ±15%, ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%,±2%, or ±1% about a reference quantity, level, value, number, frequency,percentage, dimension, size, amount, weight or length.

It will be noted that the structure of some of the compounds of theapplication include asymmetric (chiral) carbon or sulfur atoms. It is tobe understood accordingly that the isomers arising from such asymmetryare included herein, unless indicated otherwise. Such isomers can beobtained in substantially pure form by classical separation techniquesand by stereochemically controlled synthesis. The compounds of thisapplication may exist in stereoisomeric form, therefore can be producedas individual stereoisomers or as mixtures.

The term “isomerism” means compounds that have identical molecularformulae but that differ in the nature or the sequence of bonding oftheir atoms or in the arrangement of their atoms in space. Isomers thatdiffer in the arrangement of their atoms in space are termed“stereoisomers”. Stereoisomers that are not mirror images of one anotherare termed “diastereoisomers”, and stereoisomers that arenon-superimposable mirror images are termed “enantiomers”, or sometimesoptical isomers. A carbon atom bonded to four nonidentical substituentsis termed a “chiral center” whereas a sulfur bound to three or fourdifferent substitutents, e.g. sulfoxides or sulfinimides, is likewisetermed a “chiral center”.

The term “chiral isomer” means a compound with at least one chiralcenter. It has two enantiomeric forms of opposite chirality and mayexist either as an individual enantiomer or as a mixture of enantiomers.A mixture containing equal amounts of individual enantiomeric forms ofopposite chirality is termed a “racemic mixture”. A compound that hasmore than one chiral center has 2n-1 enantiomeric pairs, where n is thenumber of chiral centers. Compounds with more than one chiral center mayexist as either an individual diastereomer or as a mixture ofdiastereomers, termed a “diastereomeric mixture”. When one chiral centeris present, a stereoisomer may be characterized by the absoluteconfiguration (R or S) of that chiral center. Alternatively, when one ormore chiral centers are present, a stereoisomer may be characterized as(+) or (−). Absolute configuration refers to the arrangement in space ofthe substituents attached to the chiral center. The substituentsattached to the chiral center under consideration are ranked inaccordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn etal, Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahn et al.,Angew. Chem. 1966, 78, 413; Cahn and Ingold, J Chem. Soc. 1951 (London),612; Cahn et al., Experientia 1956, 12, 81; Cahn, J., Chem. Educ. 1964,41, 116).

The term “geometric Isomers” means the diastereomers that owe theirexistence to hindered rotation about double bonds. These configurationsare differentiated in their names by the prefixes cis and trans, or Zand E, which indicate that the groups are on the same or opposite sideof the double bond in the molecule according to the Cahn-Ingold-Prelogrules. Further, the structures and other compounds discussed in thisapplication include all atropic isomers thereof.

The term “atropic isomers” are a type of stereoisomer in which the atomsof two isomers are arranged differently in space. Atropic isomers owetheir existence to a restricted rotation caused by hindrance of rotationof large groups about a central bond. Such atropic isomers typicallyexist as a mixture, however as a result of recent advances inchromatography techniques, it has been possible to separate mixtures oftwo atropic isomers in select cases.

The terms “crystal polymorphs” or “polymorphs” or “crystal forms” meanscrystal structures in which a compound (or salt or solvate thereof) cancrystallize in different crystal packing arrangements, all of which havethe same elemental composition. Different crystal forms usually havedifferent X-ray diffraction patterns, infrared spectral, melting points,density hardness, crystal shape, optical and electrical properties,stability and solubility. Recrystallization solvent, rate ofcrystallization, storage temperature, and other factors may cause onecrystal form to dominate. Crystal polymorphs of the compounds can beprepared by crystallization under different conditions.

The term “derivative” refers to compounds that have a common corestructure, and are substituted with various groups as described herein.

The term “bioisostere” refers to a compound resulting from the exchangeof an atom or of a group of atoms with another, broadly similar, atom orgroup of atoms. The objective of a bioisosteric replacement is to createa new compound with similar biological properties to the parentcompound. The bioisosteric replacement may be physicochemically ortopologically based. Examples of carboxylic acid bioisosteres includeacyl sulfonimides, tetrazoles, sulfonates, and phosphonates. See, e.g.,Patani and LaVoie, Chem. Rev. 96, 3147-3176 (1996).

The phrases “parenteral administration” and “administered parenterally”are art-recognized terms, and include modes of administration other thanenteral and topical administration, such as injections, and include,without limitation, intravenous, intramuscular, intrapleural,intravascular, intrapericardial, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular,subarachnoid, intraspinal and intrastemal injection and infusion.

The term “treating” is art-recognized and includes inhibiting a disease,disorder or condition in a subject, e.g., impeding its progress; andrelieving the disease, disorder or condition, e.g., causing regressionof the disease, disorder and/or condition. Treating the disease orcondition includes ameliorating at least one symptom of the particulardisease or condition, even if the underlying pathophysiology is notaffected.

The term “preventing” is art-recognized and includes stopping a disease,disorder or condition from occurring in a subject, which may bepredisposed to the disease, disorder and/or condition but has not yetbeen diagnosed as having it. Preventing a condition related to a diseaseincludes stopping the condition from occurring after the disease hasbeen diagnosed but before the condition has been diagnosed.

The term “pharmaceutical composition” refers to a formulation containingthe disclosed compounds in a form suitable for administration to asubject. In a preferred embodiment, the pharmaceutical composition is inbulk or in unit dosage form. The unit dosage form is any of a variety offorms, including, for example, a capsule, an IV bag, a tablet, a singlepump on an aerosol inhaler, or a vial. The quantity of active ingredient(e.g., a formulation of the disclosed compound or salts thereof) in aunit dose of composition is an effective amount and is varied accordingto the particular treatment involved. One skilled in the art willappreciate that it is sometimes necessary to make routine variations tothe dosage depending on the age and condition of the patient. The dosagewill also depend on the route of administration. A variety of routes arecontemplated, including oral, pulmonary, rectal, parenteral,transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal,intranasal, inhalational, and the like. Dosage forms for the topical ortransdermal administration of a compound described herein includespowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches, nebulized compounds, and inhalants. In a preferred embodiment,the active compound is mixed under sterile conditions with apharmaceutically acceptable carrier, and with any preservatives,buffers, or propellants that are required.

The term “flash dose” refers to compound formulations that are rapidlydispersing dosage forms.

The term “immediate release” is defined as a release of compound from adosage form in a relatively brief period of time, generally up to about60 minutes. The term “modified release” is defined to include delayedrelease, extended release, and pulsed release. The term “pulsed release”is defined as a series of releases of drug from a dosage form. The term“sustained release” or “extended release” is defined as continuousrelease of a compound from a dosage form over a prolonged period.

The phrase “pharmaceutically acceptable” is art-recognized. In certainembodiments, the term includes compositions, polymers and othermaterials and/or dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humanbeings and animals without excessive toxicity, irritation, allergicresponse, or other problem or complication, commensurate with areasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” is art-recognized, andincludes, for example, pharmaceutically acceptable materials,compositions or vehicles, such as a liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting any subject composition from one organ, or portion of thebody, to another organ, or portion of the body. Each carrier must be“acceptable” in the sense of being compatible with the other ingredientsof a subject composition and not injurious to the patient. In certainembodiments, a pharmaceutically acceptable carrier is non-pyrogenic.Some examples of materials which may serve as pharmaceuticallyacceptable carriers include: (1) sugars, such as lactose, glucose andsucrose; (2) starches, such as corn starch and potato starch; (3)cellulose, and its derivatives, such as sodium carboxymethyl cellulose,ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5)malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter andsuppository waxes; (9) oils, such as peanut oil, cottonseed oil,sunflower oil, sesame oil, olive oil, corn oil and soybean oil; (10)glycols, such as propylene glycol; (11) polyols, such as glycerin,sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyloleate and ethyl laurate; (13) agar; (14) buffering agents, such asmagnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19)ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxiccompatible substances employed in pharmaceutical formulations.

The compounds of the application are capable of further forming salts.All of these forms are also contemplated herein.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. For example, the saltcan be an acid addition salt. One embodiment of an acid addition salt isa hydrochloride salt. The pharmaceutically acceptable salts can besynthesized from a parent compound that contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, non-aqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrilebeing preferred. Lists of salts are found in Remington's PharmaceuticalSciences, 18th ed. (Mack Publishing Company, 1990).

The compounds described herein can also be prepared as esters, forexample pharmaceutically acceptable esters. For example, a carboxylicacid function group in a compound can be converted to its correspondingester, e.g., a methyl, ethyl, or other ester. Also, an alcohol group ina compound can be converted to its corresponding ester, e.g., anacetate, propionate, or other ester.

The compounds described herein can also be prepared as prodrugs, forexample pharmaceutically acceptable prodrugs. The terms “pro-drug” and“prodrug” are used interchangeably herein and refer to any compound,which releases an active parent drug in vivo. Since prodrugs are knownto enhance numerous desirable qualities of pharmaceuticals (e.g.,solubility, bioavailability, manufacturing, etc.) the compounds can bedelivered in prodrug form. Thus, the compounds described herein areintended to cover prodrugs of the presently claimed compounds, methodsof delivering the same and compositions containing the same. “Prodrugs”are intended to include any covalently bonded carriers that release anactive parent drug in vivo when such prodrug is administered to asubject. Prodrugs are prepared by modifying functional groups present inthe compound in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent compound. Prodrugsinclude compounds wherein a hydroxy, amino, sulfhydryl, carboxy, orcarbonyl group is bonded to any group that may be cleaved in vivo toform a free hydroxyl, free amino, free sulfhydryl, free carboxy or freecarbonyl group, respectively. Prodrugs can also include a precursor(forerunner) of a compound described herein that undergoes chemicalconversion by metabolic processes before becoming an active or moreactive pharmacological agent or active compound described herein.

Examples of prodrugs include, but are not limited to, esters (e.g.,acetate, dialkylaminoacetates, formates, phosphates, sulfates, andbenzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl)of hydroxy functional groups, ester groups (e.g., ethyl esters,morpholinoethanol esters) of carboxyl functional groups, N-acylderivatives (e.g., N-acetyl)N-Mannich bases, Schiff bases and enaminonesof amino functional groups, oximes, acetals, ketals and enol esters ofketone and aldehyde functional groups in compounds, and the like, aswell as sulfides that are oxidized to form sulfoxides or sulfones.

The term “protecting group” refers to a grouping of atoms that whenattached to a reactive group in a molecule masks, reduces or preventsthat reactivity. Examples of protecting groups can be found in Green andWuts, Protective Groups in Organic Chemistry, (Wiley, 2.sup.nd ed.1991); Harrison and Harrison et al., Compendium of Synthetic OrganicMethods, Vols. 1-8 (John Wiley and Sons, 1971-1996); and Kocienski,Protecting Groups, (Verlag, 3^(rd) ed. 2003).

The term “amine protecting group” is intended to mean a functional groupthat converts an amine, amide, or other nitrogen-containing moiety intoa different chemical group that is substantially inert to the conditionsof a particular chemical reaction. Amine protecting groups arepreferably removed easily and selectively in good yield under conditionsthat do not affect other functional groups of the molecule. Examples ofamine protecting groups include, but are not limited to, formyl, acetyl,benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, t-butyloxycarbonyl(Boc), p-methoxybenzyl, methoxymethyl, tosyl, trifluoroacetyl,trimethylsilyl (TMS), fluorenyl-methyloxycarbonyl,2-trimethylsilyl-ethyoxycarbonyl, 1-methyl-1-(4-biphenylyl)ethoxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl (CBZ),2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted tritylgroups, 9-fluorenylmethyloxycarbonyl (FMOC), nitro-veratryloxycarbonyl(NVOC), and the like. Those of skill in the art can identify othersuitable amine protecting groups.

Representative hydroxy protecting groups include those where the hydroxygroup is either acylated or alkylated such as benzyl, and trityl ethersas well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethersand allyl ethers.

Additionally, the salts of the compounds described herein, can exist ineither hydrated or unhydrated (the anhydrous) form or as solvates withother solvent molecules. Non-limiting examples of hydrates includemonohydrates, dihydrates, etc. Nonlimiting examples of solvates includeethanol solvates, acetone solvates, etc.

The term “solvates” means solvent addition forms that contain eitherstoichiometric or non-stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate, when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrate.

The compounds, salts and prodrugs described herein can exist in severaltautomeric forms, including the enol and imine form, and the keto andenamine form and geometric isomers and mixtures thereof. Tautomers existas mixtures of a tautomeric set in solution. In solid form, usually onetautomer predominates. Even though one tautomer may be described, thepresent application includes all tautomers of the present compounds. Atautomer is one of two or more structural isomers that exist inequilibrium and are readily converted from one isomeric form to another.This reaction results in the formal migration of a hydrogen atomaccompanied by a switch of adjacent conjugated double bonds. Insolutions where tautomerization is possible, a chemical equilibrium ofthe tautomers will be reached. The exact ratio of the tautomers dependson several factors, including temperature, solvent, and pH. The conceptof tautomers that are interconvertable by tautomerizations is calledtautomerism.

Of the various types of tautomerism that are possible, two are commonlyobserved. In keto-enol tautomerism a simultaneous shift of electrons anda hydrogen atom occurs.

Tautomerizations can be catalyzed by: Base: 1. deprotonation; 2.formation of a delocalized anion (e.g., an enolate); 3. protonation at adifferent position of the anion; Acid: 1. protonation; 2. formation of adelocalized cation; 3. deprotonation at a different position adjacent tothe cation.

The term “analogue” refers to a chemical compound that is structurallysimilar to another but differs slightly in composition (as in thereplacement of one atom by an atom of a different element or in thepresence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analogue is acompound that is similar or comparable in function and appearance, butnot in structure or origin to the reference compound.

A “patient,” “subject,” or “host” to be treated by the subject methodmay mean either a human or non-human animal, such as a mammal, a fish, abird, a reptile, or an amphibian. Thus, the subject of the hereindisclosed methods can be a human, non-human primate, horse, pig, rabbit,dog, sheep, goat, cow, cat, guinea pig or rodent. The term does notdenote a particular age or sex. Thus, adult and newborn subjects, aswell as fetuses, whether male or female, are intended to be covered. Inone aspect, the subject is a mammal. A patient refers to a subjectafflicted with a disease or disorder.

The terms “prophylactic” or “therapeutic” treatment is art-recognizedand includes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic, i.e., it protects thehost against developing the unwanted condition, whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The terms “therapeutic agent”, “drug”, “medicament” and “bioactivesubstance” are art-recognized and include molecules and other agentsthat are biologically, physiologically, or pharmacologically activesubstances that act locally or systemically in a patient or subject totreat a disease or condition. The terms include without limitationpharmaceutically acceptable salts thereof and prodrugs. Such agents maybe acidic, basic, or salts; they may be neutral molecules, polarmolecules, or molecular complexes capable of hydrogen bonding; they maybe prodrugs in the form of ethers, esters, amides and the like that arebiologically activated when administered into a patient or subject.

The phrase “therapeutically effective amount” or “pharmaceuticallyeffective amount” is an art-recognized term. In certain embodiments, theterm refers to an amount of a therapeutic agent that produces somedesired effect at a reasonable benefit/risk ratio applicable to anymedical treatment. In certain embodiments, the term refers to thatamount necessary or sufficient to eliminate, reduce or maintain a targetof a particular therapeutic regimen. The effective amount may varydepending on such factors as the disease or condition being treated, theparticular targeted constructs being administered, the size of thesubject or the severity of the disease or condition. One of ordinaryskill in the art may empirically determine the effective amount of aparticular compound without necessitating undue experimentation. Incertain embodiments, a therapeutically effective amount of a therapeuticagent for in vivo use will likely depend on a number of factors,including: the rate of release of an agent from a polymer matrix, whichwill depend in part on the chemical and physical characteristics of thepolymer; the identity of the agent; the mode and method ofadministration; and any other materials incorporated in the polymermatrix in addition to the agent.

The term “ED50” is art-recognized. In certain embodiments, ED50 meansthe dose of a drug, which produces 50% of its maximum response oreffect, or alternatively, the dose, which produces a pre-determinedresponse in 50% of test subjects or preparations. The term “LD50” isart-recognized. In certain embodiments, LD50 means the dose of a drug,which is lethal in 50% of test subjects. The term “therapeutic index” isan art-recognized term, which refers to the therapeutic index of a drug,defined as LD50/ED50.

The terms “IC₅₀,” or “half maximal inhibitory concentration” is intendedto refer to the concentration of a substance (e.g., a compound or adrug) that is required for 50% inhibition of a biological process, orcomponent of a process, including a protein, subunit, organelle,ribonucleoprotein, etc.

With respect to any chemical compounds, the present application isintended to include all isotopes of atoms occurring in the presentcompounds. Isotopes include those atoms having the same atomic numberbut different mass numbers. By way of general example and withoutlimitation, isotopes of hydrogen include tritium and deuterium, andisotopes of carbon include C-13 and C-14.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent can be bonded to any atom in thering. When a substituent is listed without indicating the atom via whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent can be bonded via any atom in suchsubstituent. Combinations of substituents and/or variables arepermissible, but only if such combinations result in stable compounds.

When an atom or a chemical moiety is followed by a subscripted numericrange (e.g., C₁₋₆), it is meant to encompass each number within therange as well as all intermediate ranges. For example, “C₁₋₆ alkyl” ismeant to include alkyl groups with 1, 2, 3, 4, 5, 6, 1-6, 1-5, 1-4, 1-3,1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, and 5-6 carbons.

The term “alkyl” is intended to include both branched (e.g., isopropyl,tert-butyl, isobutyl), straight-chain e.g., methyl, ethyl, propyl,butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl), and cycloalkyl(e.g., alicyclic) groups (e.g., cyclopropyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl), alkyl substituted cycloalkyl groups, andcycloalkyl substituted alkyl groups. Such aliphatic hydrocarbon groupshave a specified number of carbon atoms. For example, C₁₋₆ alkyl isintended to include C₁, C₂, C₃, C₄, C₅, and C₆ alkyl groups. As usedherein, “lower alkyl” refers to alkyl groups having from 1 to 6 carbonatoms in the backbone of the carbon chain. “Alkyl” further includesalkyl groups that have oxygen, nitrogen, sulfur or phosphorous atomsreplacing one or more hydrocarbon backbone carbon atoms. In certainembodiments, a straight chain or branched chain alkyl has six or fewercarbon atoms in its backbone (e.g., C₁-C₆ for straight chain, C₃-C₆ forbranched chain), for example four or fewer. Likewise, certaincycloalkyls have from three to eight carbon atoms in their ringstructure, such as five or six carbons in the ring structure.

The term “substituted alkyls” refers to alkyl moieties havingsubstituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example, alkyl,alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Cycloalkyls can be further substituted, e.g.,with the substituents described above. An “alkylaryl” or an “aralkyl”moiety is an alkyl substituted with an aryl (e.g., phenylmethyl(benzyl)). If not otherwise indicated, the terms “alkyl” and “loweralkyl” include linear, branched, cyclic, unsubstituted, substituted,and/or heteroatom-containing alkyl or lower alkyl, respectively.

The term “alkenyl” refers to a linear, branched or cyclic hydrocarbongroup of 2 to about 24 carbon atoms containing at least one double bond,such as ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl,octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl,cyclopentenyl, cyclohexenyl, cyclooctenyl, and the like. Generally,although again not necessarily, alkenyl groups can contain 2 to about 18carbon atoms, and more particularly 2 to 12 carbon atoms. The term“lower alkenyl” refers to an alkenyl group of 2 to 6 carbon atoms, andthe specific term “cycloalkenyl” intends a cyclic alkenyl group,preferably having 5 to 8 carbon atoms. The term “substituted alkenyl”refers to alkenyl substituted with one or more substituent groups, andthe terms “heteroatom-containing alkenyl” and “heteroalkenyl” refer toalkenyl or heterocycloalkenyl (e.g., heterocylcohexenyl) in which atleast one carbon atom is replaced with a heteroatom. If not otherwiseindicated, the terms “alkenyl” and “lower alkenyl” include linear,branched, cyclic, unsubstituted, substituted, and/orheteroatom-containing alkenyl and lower alkenyl, respectively.

The term “alkynyl” refers to a linear or branched hydrocarbon group of 2to 24 carbon atoms containing at least one triple bond, such as ethynyl,n-propynyl, and the like. Generally, although again not necessarily,alkynyl groups can contain 2 to about 18 carbon atoms, and moreparticularly can contain 2 to 12 carbon atoms. The term “lower alkynyl”intends an alkynyl group of 2 to 6 carbon atoms. The term “substitutedalkynyl” refers to alkynyl substituted with one or more substituentgroups, and the terms “heteroatom-containing alkynyl” and“heteroalkynyl” refer to alkynyl in which at least one carbon atom isreplaced with a heteroatom. If not otherwise indicated, the terms“alkynyl” and “lower alkynyl” include linear, branched, unsubstituted,substituted, and/or heteroatom-containing alkynyl and lower alkynyl,respectively.

The terms “alkyl”, “alkenyl”, and “alkynyl” are intended to includemoieties which are diradicals, i.e., having two points of attachment. Anonlimiting example of such an alkyl moiety that is a diradical is—CH₂CH₂—, i.e., a C₂ alkyl group that is covalently bonded via eachterminal carbon atom to the remainder of the molecule.

The term “alkoxy” refers to an alkyl group bound through a single,terminal ether linkage; that is, an “alkoxy” group may be represented as—O-alkyl where alkyl is as defined above. A “lower alkoxy” group intendsan alkoxy group containing 1 to 6 carbon atoms, and includes, forexample, methoxy, ethoxy, n-propoxy, isopropoxy, t-butyloxy, etc.Preferred substituents identified as “C₁-C₆ alkoxy” or “lower alkoxy”herein contain 1 to 3 carbon atoms, and particularly preferred suchsubstituents contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy).

The term “aryl” refers to an aromatic substituent containing a singlearomatic ring or multiple aromatic rings that are fused together,directly linked, or indirectly linked (such that the different aromaticrings are bound to a common group such as a methylene or ethylenemoiety). Aryl groups can contain 5 to 20 carbon atoms, and particularlypreferred aryl groups can contain 5 to 14 carbon atoms. Examples of arylgroups include benzene, phenyl, pyrrole, furan, thiophene, thiazole,isothiazole, imidazole, triazole, tetrazole, pyrazole, oxazole,isooxazole, pyridine, pyrazine, pyridazine, and pyrimidine, and thelike. Furthermore, the term “aryl” includes multicyclic aryl groups,e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole,benzodioxazole, benzothiazole, benzoimidazole, benzothiophene,methylenedioxyphenyl, quinoline, isoquinoline, napthridine, indole,benzofuran, purine, benzofuran, deazapurine, or indolizine. Those arylgroups having heteroatoms in the ring structure may also be referred toas “aryl heterocycles”, “heterocycles,” “heteroaryls” or“heteroaromatics”. The aromatic ring can be substituted at one or morering positions with such substituents as described above, as forexample, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl,alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diaryl amino, and al kylaryl amino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Aryl groups can also be fused or bridged withalicyclic or heterocyclic rings, which are not aromatic so as to form amulticyclic system (e.g., tetralin, methylenedioxyphenyl). If nototherwise indicated, the term “aryl” includes unsubstituted,substituted, and/or heteroatom-containing aromatic substituents.

The term “alkaryl” refers to an aryl group with an alkyl substituent,and the term “aralkyl” refers to an alkyl group with an arylsubstituent, wherein “aryl” and “alkyl” are as defined above. Exemplaryaralkyl groups contain 6 to 24 carbon atoms, and particularly preferredaralkyl groups contain 6 to 16 carbon atoms. Examples of aralkyl groupsinclude, without limitation, benzyl, 2-phenyl-ethyl, 3-phenyl-propyl,4-phenyl-butyl, 5-phenyl-pentyl, 4-phenylcyclohexyl, 4-benzylcyclohexyl,4-phenylcyclohexylmethyl, 4-benzylcyclohexylmethyl, and the like.Alkaryl groups include, for example, p-methylphenyl, 2,4-dimethylphenyl,p-cyclohexylphenyl, 2,7-dimethylnaphthyl, 7-cyclooctylnaphthyl,3-ethyl-cyclopenta-1,4-diene, and the like.

The terms “heterocyclyl” or “heterocyclic group” include closed ringstructures, e.g., 3- to 10-, or 4- to 7-membered rings, which includeone or more heteroatoms. “Heteroatom” includes atoms of any elementother than carbon or hydrogen. Examples of heteroatoms include nitrogen,oxygen, sulfur and phosphorus.

Heterocyclyl groups can be saturated or unsaturated and includepyrrolidine, oxolane, thiolane, piperidine, piperazine, morpholine,lactones, lactams, such as azetidinones and pyrrolidinones, sultams, andsultones. Heterocyclic groups such as pyrrole and furan can havearomatic character. They include fused ring structures, such asquinoline and isoquinoline. Other examples of heterocyclic groupsinclude pyridine and purine. The heterocyclic ring can be substituted atone or more positions with such substituents as described above, as forexample, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl,cyano, azido, heterocyclyl, or an aromatic or heteroaromatic moiety.Heterocyclic groups can also be substituted at one or more constituentatoms with, for example, a lower alkyl, a lower alkenyl, a lower alkoxy,a lower alkylthio, a lower alkylamino, a lower alkylcarboxyl, a nitro, ahydroxyl, —CF₃, or —CN, or the like.

The term “halo” or “halogen” refers to fluoro, chloro, bromo, and iodo.“Counterion” is used to represent a small, negatively charged speciessuch as fluoride, chloride, bromide, iodide, hydroxide, acetate, andsulfate. The term sulfoxide refers to a sulfur attached to 2 differentcarbon atoms and one oxygen and the S—O bond can be graphicallyrepresented with a double bond (S═O), a single bond without charges(S—O) or a single bond with charges [S(+)—O(−)].

The terms “substituted” as in “substituted alkyl,” “substituted aryl,”and the like, as alluded to in some of the aforementioned definitions,is meant that in the alkyl, aryl, or other moiety, at least one hydrogenatom bound to a carbon (or other) atom is replaced with one or morenon-hydrogen substituents. Examples of such substituents include,without limitation: functional groups such as halo, hydroxyl, silyl,sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀aryloxy, acyl (including C₂-C₂₄ alkylcarbonyl (—CO-alkyl) and C₆-C₂₀arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C₂-C₂₄ alkoxycarbonyl(—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl (—(CO)—O-aryl), C₂-C₂₄alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀ arylcarbonato (—O—(CO)—O-aryl),carboxy (—COOH), carboxylato (—COO—), carbamoyl (—(CO)—NH₂),mono-(C₁-C₂₄ alkyl)-substituted carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),di-(C₁-C₄ alkyl)-substituted carbamoyl (—(CO)—N(C₁-C₂₄ alkyl)₂),mono-substituted arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl(—(CS)—NH₂), carbamido (—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻),cyanato (—O—CN), isocyanato (—ON⁺C⁻), isothiocyanato (—S—CN), azido(—N═N⁺═N⁻), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), mono-and di-(C₁-C₂₄ alkyl)-substituted amino, mono- and di-(C₅-C₂₀aryl)-substituted amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀arylamido (—NH—(CO)-aryl), imino (—CR═NH where R=hydrogen, C₁-C₂₄ alkyl,C₅-C₂₀ aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino(—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), phosphono (—P(O)(OH)₂), phosphonato(—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂), and phosphino(—PH₂); and the hydrocarbyl moieties C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl, and C₆-C₂₄ aralkyl.

In addition, the aforementioned functional groups may, if a particulargroup permits, be further substituted with one or more additionalfunctional groups or with one or more hydrocarbyl moieties such as thosespecifically enumerated above. Analogously, the above-mentionedhydrocarbyl moieties may be further substituted with one or morefunctional groups or additional hydrocarbyl moieties such as thosespecifically enumerated.

When the term “substituted” appears prior to a list of possiblesubstituted groups, it is intended that the term apply to every memberof that group. For example, the phrase “substituted alkyl, alkenyl, andaryl” is to be interpreted as “substituted alkyl, substituted alkenyl,and substituted aryl.” Analogously, when the term“heteroatom-containing” appears prior to a list of possibleheteroatom-containing groups, it is intended that the term apply toevery member of that group. For example, the phrase“heteroatom-containing alkyl, alkenyl, and aryl” is to be interpreted as“heteroatom-containing alkyl, substituted alkenyl, and substituted aryl.

“Optional” or “optionally” means that the subsequently describedcircumstance may or may not occur, so that the description includesinstances where the circumstance occurs and instances where it does not.For example, the phrase “optionally substituted” means that anon-hydrogen substituent may or may not be present on a given atom, and,thus, the description includes structures wherein a non-hydrogensubstituent is present and structures wherein a non-hydrogen substituentis not present.

The terms “stable compound” and “stable structure” are meant to indicatea compound that is sufficiently robust to survive isolation, and asappropriate, purification from a reaction mixture, and formulation intoan efficacious therapeutic agent.

The terms “free compound” is used herein to describe a compound in theunbound state.

Throughout the description, where compositions are described as having,including, or comprising, specific components, it is contemplated thatcompositions also consist essentially of, or consist of, the recitedcomponents. Similarly, where methods or processes are described ashaving, including, or comprising specific process steps, the processesalso consist essentially of, or consist of, the recited processingsteps. Further, it should be understood that the order of steps or orderfor performing certain actions is immaterial so long as the compositionsand methods described herein remains operable. Moreover, two or moresteps or actions can be conducted simultaneously.

The term “small molecule” is an art-recognized term. In certainembodiments, this term refers to a molecule, which has a molecularweight of less than about 2000 amu, or less than about 1000 amu, andeven less than about 500 amu.

All percentages and ratios used herein, unless otherwise indicated, areby weight.

The terms “gene expression” or “protein expression” includes anyinformation pertaining to the amount of gene transcript or proteinpresent in a sample, as well as information about the rate at whichgenes or proteins are produced or are accumulating or being degraded(e.g., reporter gene data, data from nuclear runoff experiments,pulse-chase data etc.). Certain kinds of data might be viewed asrelating to both gene and protein expression. For example, proteinlevels in a cell are reflective of the level of protein as well as thelevel of transcription, and such data is intended to be included by thephrase “gene or protein expression information”. Such information may begiven in the form of amounts per cell, amounts relative to a controlgene or protein, in unitless measures, etc.; the term “information” isnot to be limited to any particular means of representation and isintended to mean any representation that provides relevant information.The term “expression levels” refers to a quantity reflected in orderivable from the gene or protein expression data, whether the data isdirected to gene transcript accumulation or protein accumulation orprotein synthesis rates, etc.

The terms “healthy” and “normal” are used interchangeably herein torefer to a subject or particular cell or tissue that is devoid (at leastto the limit of detection) of a disease condition.

The term “nucleic acid” refers to polynucleotides such asdeoxyribonucleic acid (DNA), and, where appropriate, ribonucleic acid(RNA). The term should also be understood to include analogues of eitherRNA or DNA made from nucleotide analogues, and, as applicable to theembodiment being described, single-stranded (such as sense or antisense)and double-stranded polynucleotides. In some embodiments, “nucleic acid”refers to inhibitory nucleic acids. Some categories of inhibitorynucleic acid compounds include antisense nucleic acids, RNAi constructs,and catalytic nucleic acid constructs. Such categories of nucleic acidsare well-known in the art.

Embodiments described herein relate to compositions and methods formodulating hematopoietic stem cells and hematopoiesis. As described inthe Examples below, it was found that that inhibitors of short-chaindehydrogenase activity, such as 15-PGDH inhibitors, can be administeredto a subject in need thereof, cells of the subject, and/or tissue of thesubject alone or in combination with a cytokine to increase and/ormobilize hematopoietic stem cells and/or neutrophils in the blood,marrow, and/or tissue of the subject.

In some embodiments, the 15-PGDH inhibitor can be administered to thesubject or a preparation of hematopoietic stem cells, such as peripheralblood hematopoietic stem cells or umbilical cord stem cells of thesubject, to increase the fitness of the stem cell preparation as a donorgraft or to decrease the number of units of umbilical cord bloodrequired for transplantation.

Hematopoietic stem cells are multipotent stem cells that give rise toall the blood cell types of an organism, including myeloid (e.g.,monocytes and macrophages, neutrophils, basophils, eosinophils,erythrocytes, megakaryocytes/platelets, dendritic cells), and lymphoidlineages (e.g., T-cells, B-cells, NK-cells), and others known in the art(See Fei, R., et al, U.S. Pat. No. 5,635,387; McGlave, et al, U.S. Pat.No. 5,460,964; Simmons, P., et al, U.S. Pat. No. 5,677,136; Tsukamoto,et al, U.S. Pat. No. 5,750,397; Schwartz, et al, U.S. Pat. No.5,759,793; DiGuisto, et al, U.S. Pat. No. 5,681,599; Tsukamoto, et al,U.S. Pat. No. 5,716,827). Hematopoietic stem cells (HSCs) give rise tocommitted hematopoietic progenitor cells (HPCs) that are capable ofgenerating the entire repertoire of mature blood cells over the lifetimeof an organism.

Hematopoietic stem cells and hematopoietic progenitor cells aredescribed herein generally as hematopoietic stem cells unless notedotherwise and can refer to cells or populations identified by thepresence of the antigenic marker CD34 (CD34⁺). In some embodiments, thehematopoietic stem cells can be identified by the presence of theantigenic marker CD34 and the absence of lineage (lin) markers and aretherefore characterized as CD34⁺/lin⁻ cells.

The hematopoietic stem cells used in the methods described herein may beobtained from any suitable source of hematopoietic stem and progenitorcells and can be provided as a high purified population of hematopoieticstem cells or as composition that includes about 0.01% to about 100% ofhematopoietic stem cells. For example, hematopoietic stem cells may beprovided in compositions, such as unfractionated bone marrow (where thehematopoietic stem cells comprise less than about 1% of the bone marrowcell population), umbilical cord blood, placental blood, placenta, fetalblood, fetal liver, fetal spleen, Wharton's jelly, or mobilizedperipheral blood.

Suitable sources of hematopoietic stem cells can be isolated or obtainedfrom an organ of the body containing cells of hematopoietic origin. Theisolated cells can include cells that are removed from their originalenvironment. For example, a cell is isolated if it is separated fromsome or all of the components that normally accompany it in its nativestate. For example, an “isolated population of cells,” an “isolatedsource of cells,” or “isolated hematopoietic stem cells” and the like,as used herein, refer to in vitro or ex vivo separation of one or morecells from their natural cellular environment, and from association withother components of the tissue or organ, i.e., it is not significantlyassociated with in vivo substances.

Hematopoietic stem cells can be obtained or isolated from bone marrow ofadults, which includes femurs, hip, ribs, sternum, and other bones. Bonemarrow aspirates containing hematopoietic stem cells can be obtained orisolated directly from the hip using a needle and syringe. Other sourcesof hematopoietic stem cells include umbilical cord blood, placentalblood, mobilized peripheral blood, Wharton's jelly, placenta, fetalblood, fetal liver, or fetal spleen. In particular embodiments,harvesting a sufficient quantity of hematopoietic stem cells for use intherapeutic applications may require mobilizing the stem and progenitorcells in the donor.

“Hematopoietic stem cell mobilization” refers to the release of stemcells from the bone marrow into the peripheral blood circulation for thepurpose of leukapheresis, prior to stem cell transplantation. Byincreasing the number of stem cells harvested from the donor, the numberof stem cells available for therapeutic applications can besignificantly improved. Hematopoietic growth factors, e.g., granulocytecolony stimulating factor (G-CSF) or chemotherapeutic agents often areused to stimulate the mobilization. Commercial cell mobilization agentsexist and can be used in combination with G-CSF to mobilize sufficientquantities of hematopoietic stem and progenitor cells fortransplantation into a subject. For example, G-CSF and MOZOBIL(Plerixafor, Genzyme Corporation) can be administered to a donor inorder to harvest a sufficient number of hematopoietic cells fortransplantation. Other methods of mobilizing hematopoietic stem cellswould be apparent to one having skill in the art.

In some embodiments, hematopoietic stem cells are obtained fromumbilical cord blood. Cord blood can be harvested according totechniques known in the art (see, e.g., U.S. Pat. Nos. 7,147,626 and7,131,958, herein incorporated by reference for such methodologies).

In one embodiment, hematopoietic stem cells can be obtained frompluripotent stem cell sources, e.g., induced pluripotent stem cells(iPSCs) and embryonic stem cells (ESCs). As used herein, the term“induced pluripotent stem cell” or “iPSC” refers to a non-pluripotentcell that has been reprogrammed to a pluripotent state. Once the cellsof a subject have been reprogrammed to a pluripotent state, the cellscan then be programmed to a desired cell type, such as a hematopoieticstem or progenitor cell. As used herein, the term “reprogramming” refersto a method of increasing the potency of a cell to a less differentiatedstate. As used herein, the term “programming” refers to a method ofdecreasing the potency of a cell or differentiating the cell to a moredifferentiated state.

In some embodiments, the 15-PGDH inhibitor can be administered to thesubject or to hematopoietic stem cells, which can be administered orcontacted ex vivo with one or more 15-PGDH inhibitors described hereinto provide a therapeutic composition. In one embodiment, the therapeuticcompositions or preparations can include the 15-PGDH inhibitor or apopulation of hematopoietic stem cells treated ex vivo with a one ormore 15-PGDH inhibitor. In certain embodiments, the therapeuticcomposition including the enhanced hematopoietic stem cells can includewhole bone marrow, umbilical cord blood, or mobilized peripheral blood.

In particular embodiments, the therapeutic composition includes apopulation of cells, wherein the population of cells is about 95% toabout 100% hematopoietic stem cells. The use of therapeutic compositionsof highly purified hematopoietic stem cells, e.g., a compositioncomprising a population of cells wherein the cells comprise about 95%hematopoietic stem cells, may improve the efficiency of stem celltherapies. Currently practiced methods of transplantations typically useunfractionated mixtures of cells where hematopoietic stem cells compriseless than 1% of the total cell population.

In some embodiments, the therapeutic composition comprises a populationof cells, wherein the population of cells comprises less than about0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, or 30% hematopoietic stemcells. The population of cells in other embodiments comprises more thanabout 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, or 30% hematopoieticstem cells. In still other embodiments, the population of cells is about0.1% to about 1%, about 1% to about 3%, about 3% to about 5%, about10%-15%, about 15%-20%, about 20%-25%, about 25%-30%, about 30%-35%,about 35%-40%, about 40%-45%, about 45%-50%, about 60%-70%, about70%-80%, about 80%-90%, about 90%-95%, or about 95% to about 100%hematopoietic stem cells.

Hematopoietic stem cells in the therapeutic compositions describedherein can be autologous/autogeneic (“self) or non-autologous(“non-self,” e.g., allogeneic, syngeneic or xenogeneic) relative to asubject to which the therapeutic composition is to be administered.“Autologous,” as used herein, refers to cells from the same subject.“Allogeneic,” as used herein, refers to cells of the same species thatdiffer genetically to the cell in comparison. “Syngeneic,” as usedherein, refers to cells of a different subject that are geneticallyidentical to the cell in comparison. “Xenogeneic,” as used herein,refers to cells of a different species to the cell in comparison.

Hematopoietic stem cells for use in the methods described herein may bedepleted of mature hematopoietic cells, such as T cells, B cells, NKcells, dendritic cells, monocytes, granulocytes, erythroid cells, andtheir committed precursors from bone marrow aspirate, umbilical cordblood, or mobilized peripheral blood (mobilized leukapheresis product).Mature, lineage committed cells are depleted by immunodepletion, forexample, by labeling solid substrates with antibodies that bind to apanel of so-called “lineage” antigens: CD2, CD3, CD11b, CD14, CD15,CD16, CD79, CD56, CD123, and CD235a. A subsequent step can be performedto further purify the population of cells, in which a substrate labeledwith antibodies that bind to the CD34⁺ antigen are used to isolateprimitive hematopoietic stem cells. Kits are commercially available forpurifying stem and progenitor cells from various cell sources and inparticular embodiments, these kits are suitable for use with the methodsdescribed herein.

In one embodiment, the amount of hematopoietic stem cells in thetherapeutic composition is at least 0.1×10⁵ cells, at least 0.5×10⁵cells, at least 1×10⁵ cells, at least 5×10⁵ cells, at least 10×10⁵cells, at least 0.5×10⁶ cells, at least 0.75×10⁶ cells, at least 1×10⁶cells, at least 1.25×10⁶ cells, at least 1.5×10⁶ cells, at least1.75×10⁶ cells, at least 2×10⁶ cells, at least 2.5×10⁶ cells, at least3×10⁶ cells, at least 4×10⁶ cells, at least 5×10⁶ cells, at least 10×10⁶cells, at least 15×10⁶ cells, at least 20×10⁶ cells, at least 25×10⁶cells, or at least 30×10⁶ cells.

In one embodiment, the amount of hematopoietic stem cells in thetherapeutic composition is the amount of hematopoietic stem cells in apartial or single cord of blood, or is at least 0.1×10⁵ cells/kg ofbodyweight, at least 0.5×10⁵ cells/kg of bodyweight, at least 1×10⁵cells/kg of bodyweight, at least 5×10⁵ cells/kg of bodyweight, at least10×10⁵ cells/kg of bodyweight, at least 0.5×10⁶ cells/kg of bodyweight,at least 0.75×10⁶ cells/kg of bodyweight, at least 1×10⁶ cells/kg ofbodyweight, at least 1.25×10⁶ cells/kg of bodyweight, at least 1.5×10⁶cells/kg of bodyweight, at least 1.75×10⁶ cells/kg of bodyweight, atleast 2×10⁶ cells/kg of bodyweight, at least 2.5×10⁶ cells/kg ofbodyweight, at least 3×10⁶ cells/kg of bodyweight, at least 4×10⁶cells/kg of bodyweight, at least 5×10⁶ cells/kg of bodyweight, at least10×10⁶ cells/kg of bodyweight, at least 15×10⁶ cells/kg of bodyweight,at least 20×10⁶ cells/kg of bodyweight, at least 25×10⁶ cells/kg ofbodyweight, or at least 30×10⁶ cells/kg of bodyweight.

Preparations of the 15-PGDH inhibitor as well as hematopoietic stemcells administered one or more 15-PGDH inhibitors and/or therapeuticcompositions that include hematopoietic stem cells and one or more15-PGDH inhibitor can be used for improving hematopoietic stem celltransplants and in treating ischemia or ischemia-damaged tissue, and inreducing further damage to ischemic tissue and/or repairing damage toischemic tissue through cell recruitment, improving vascularization inischemic tissue, improving tissue regeneration at sites of ischemia,decreasing ischemic tissue necrosis or apoptosis, and/or increasing cellsurvival at sites of ischemia. In particular embodiments, thepreparations of 15-PGDH inhibitor, 15-PGDH inhibitor treatedhematopoietic stem cells, and/or therapeutic compositions of 15-PGDHinhibitors and hematopoietic stem cells are useful to subjects in needof hematopoietic reconstitution, such as subjects that have undergone orare scheduled to undergo myeloablative therapy.

Subjects, who can be treated with the preparations of 15-PGDH inhibitor,15-PGDH inhibitor treated hematopoietic stem cells and/or therapeuticcompositions of 15-PGDH inhibitors and hematopoietic stem cells, caninclude subjects that have or that have been diagnosed with varioustypes of leukemias, anemias, lymphomas, myelomas, immune deficiencydisorders, and solid tumors. A subject also includes a human who is acandidate for stem cell transplant or bone marrow transplantation, suchas during the course of treatment for a malignant disease or a componentof gene therapy. Subjects may also include individuals or animals thatdonate stem cells or bone marrow for allogeneic transplantation. Incertain embodiments, a subject may have undergone myeloablativeirradiation therapy or chemotherapy, or may have experienced an acuteradiation or chemical insult resulting in myeloablation. In certainembodiments, a subject may have undergone irradiation therapy orchemotherapy, such as during various cancer treatments. Typical subjectsinclude animals that exhibit aberrant amounts (lower or higher amountsthan a “normal” or “healthy” subject) of one or more physiologicalactivities that can be modulated by an agent or a stem cell or marrowtransplant.

Subjects, which can be treated with the preparations of 15-PGDHinhibitors, 15-PGDH inhibitor treated hematopoietic stem cells and/ortherapeutic compositions of 15-PGDH inhibitors and hematopoietic stemcells, can also include subjects undergoing chemotherapy or radiationtherapy for cancer, as well as subjects suffering from (e.g., afflictedwith) non malignant blood disorders, particularly immunodeficiencies(e.g. SCID, Fanconi's anemia, severe aplastic anemia, or congenitalhemoglobinopathies, or metabolic storage diseases, such as Hurler'sdisease, Hunter's disease, mannosidosis, among others) or cancer,particularly hematological malignancies, such as acute leukemia, chronicleukemia (myeloid or lymphoid), lymphoma (Hodgkin's or non-Hodgkin's),multiple myeloma, myelodysplastic syndrome, or non-hematologicalcancers, such as solid tumors (including breast cancer, ovarian cancer,brain cancer, prostate cancer, lung cancer, colon cancer, skin cancer,liver cancer, or pancreatic cancer).

Subjects may also include subjects suffering from aplastic anemia, animmune disorder (severe combined immune deficiency syndrome or lupus),myelodysplasia, thalassemaia, sickle-cell disease or Wiskott-Aldrichsyndrome. In some embodiments, the subject suffers from a disorder thatis the result of an undesired side effect or complication of anotherprimary treatment, such as radiation therapy, chemotherapy, or treatmentwith a bone marrow suppressive drug, such as zidovadine, chloramphenicalor gangciclovir. Such disorders include neutropenias, anemias,thrombocytopenia, and immune dysfunction. Other subjects may havedisorders caused by an infection (e.g., viral infection, bacterialinfection or fungal infection) which causes damage to stem or progenitorcells of the bone marrow.

In addition, subjects suffering from the following conditions can alsobenefit from treatment using the preparations of 15-PGDH inhibitor,15-PGDH inhibitor treated hematopoietic stem cells and/or therapeuticcompositions of 15-PGDH inhibitors and hematopoietic stem cells:lymphocytopenia, lymphorrhea, lymphostasis, erythrocytopenia,erthrodegenerative disorders, erythroblastopenia, leukoerythroblastosis;erythroclasis, thalassemia, myelodysplasia, myelofibrosis,thrombocytopenia, disseminated intravascular coagulation (DIC), immune(autoimmune) thrombocytopenic purpura (ITP), HIV inducted ITP,myelodysplasia; thrombocytotic disease, thrombocytosis, congenitalneutropenias (such as Kostmann's syndrome and Schwachman-Diamondsyndrome), neoplastic associated neutropenias, childhood and adultcyclic neutropaenia; post-infective neutropaenia; myelodysplasticsyndrome; neutropaenia associated with chemotherapy and radiotherapy;chronic granulomatous disease; mucopolysaccharidoses; Diamond BlackfanAnemia; Sickle cell disease; or Beta thalassemia major.

In other embodiments, the preparations of 15-PGDH inhibitor, 15-PGDHinhibitor treated hematopoietic stem cells, and/or therapeuticcompositions or 15-PGDH inhibitors and hematopoietic stem cells can beused in cell-based therapy for treating ischemic tissue or treating orameliorating one or more symptoms associated with tissue ischemia,including, but not limited to, impaired, or loss of, organ function(including without limitation impairments or loss of brain, kidney, orheart function), cramping, claudication, numbness, tingling, weakness,pain, reduced wound healing, inflammation, skin discoloration, andgangrene.

In one embodiment, the subject exhibits at least one symptom of anischemic tissue or tissue damaged by ischemia. In particularembodiments, the subject is a human who is has or who is at risk ofhaving an ischemic tissue or tissue damaged by ischemia, e.g., a subjectthat has diabetes, peripheral vascular disease, thromboangiitisobliterans, vasculitis, cardiovascular disease, coronary artery diseaseor heart failure, or cerebrovascular disease, cardiovascular disease, orcerebrovascular disease.

Illustrative examples of genetic disorders, syndromic conditions,traumatic injuries, chronic conditions, medical interventions, or otherconditions that cause or are associated with ischemia, or increase therisk of ischemia in a subject, or cause a subject to exhibit more ormore symptoms of ischemia, and thus, suitable for treatment oramelioration using the methods described herein, include, but are notlimited to, acute coronary syndrome, acute lung injury (ALI), acutemyocardial infarction (AMI), acute respiratory distress syndrome (ARDS),arterial occlusive disease, arteriosclerosis, articular cartilagedefect, aseptic systemic inflammation, atherosclerotic cardiovasculardisease, autoimmune disease, bone fracture, bone fracture, brain edema,brain hypoperfusion, Buerger's disease, burns, cancer, cardiovasculardisease, cartilage damage, cerebral infarct, cerebral ischemia, cerebralstroke, cerebrovascular disease, chemotherapy-induced neuropathy,chronic infection, chronic mesenteric ischemia, claudication, congestiveheart failure, connective tissue damage, contusion, coronary arterydisease (CAD), critical limb ischemia (CLI), Crohn's disease, deep veinthrombosis, deep wound, delayed ulcer healing, delayed wound-healing,diabetes (type I and type II), diabetic neuropathy, diabetes inducedischemia, disseminated intravascular coagulation (DIC), embolic brainischemia, graft-versus-host disease, frostbite, hereditary hemorrhagictelengiectasiaischemic vascular disease, hyperoxic injury, hypoxia,inflammation, inflammatory bowel disease, inflammatory disease, injuredtendons, intermittent claudication, intestinal ischemia, ischemia,ischemic brain disease, ischemic heart disease, ischemic peripheralvascular disease, ischemic placenta, ischemic renal disease, ischemicvascular disease, ischemic-reperfusion injury, laceration, left maincoronary artery disease, limb ischemia, lower extremity ischemia,myocardial infarction, myocardial ischemia, organ ischemia,osteoarthritis, osteoporosis, osteosarcoma, Parkinson's disease,peripheral arterial disease (PAD), peripheral artery disease, peripheralischemia, peripheral neuropathy, peripheral vascular disease,pre-cancer, pulmonary edema, pulmonary embolism, remodeling disorder,renal ischemia, retinal ischemia, retinopathy, sepsis, skin ulcers,solid organ transplantation, spinal cord injury, stroke,subchondral-bone cyst, thrombosis, thrombotic brain ischemia, tissueischemia, transient ischemic attack (TIA), traumatic brain injury,ulcerative colitis, vascular disease of the kidney, vascularinflammatory conditions, von Hippel-Lindau syndrome, and wounds totissues or organs.

Other illustrative examples of genetic disorders, syndromic conditions,traumatic injuries, chronic conditions, medical interventions, or otherconditions that cause or are associated with ischemia, or increase therisk of ischemia in a subject, or cause a subject to exhibit more ormore symptoms of ischemia suitable for treatment or amelioration usingthe methods of the present invention, include, ischemia resulting fromsurgery, chemotherapy, radiation therapy, or cell, tissue, or organtransplant or graft.

In various embodiments, the methods are suitable for treatingcerebrovascular ischemia, myocardial ischemia, limb ischemia (CLI),myocardial ischemia (especially chronic myocardial ischemia), ischemiccardiomyopathy, cerebrovascular ischemia, renal ischemia, pulmonaryischemia, intestinal ischemia, and the like.

In various embodiments, the preparations or therapeutic cellcompositions disclosed herein can be used to treat an ischemic tissue inwhich it is desirable to increase the blood flow, oxygen supply, glucosesupply, or supply of nutrients to the tissue.

In some embodiments, the 15-PGDH inhibitor can be administered to apreparation of tissue stem cells, such as neural stem stems, mesenchymalstem cells, or stem cells that can generate other tissues, and/or apreparation of pluripotent stem cells.

In other embodiments, the 15-PGDH inhibitor can be administered to abone marrow graft donor or a hematopoietic stem cell donor to increasethe fitness of a donor bone marrow graft or a donor hematopoietic stemcell graft.

In other embodiments, the 15-PGDH inhibitor can also be administered tobone marrow of a subject to increase stem cells in the subject or toincrease the fitness of the marrow as a donor graft.

In yet other embodiments, the 15-PGDH inhibitor can be administered to asubject to mitigate bone marrow graft rejection, to enhance bone marrowgraft engraftment, to enhance engraftment of a hematopoietic stem cellgraft, or an umbilical cord blood stem cell graft, to enhanceengraftment of a hematopoietic stem cell graft, or an umbilical cordstem cell graft, and/or to decrease the number of units of umbilicalcord blood required for transplantation into the subject. Theadministration can be, for example, following treatment of the subjector the marrow of the subject with radiation therapy, chemotherapy, orimmunosuppressive therapy.

In other embodiments, the 15-PGDH inhibitor can be administered to arecipient of a bone marrow transplant, of a hematopoietic stem celltransplant, or of an umbilical cord blood stem cell transplant, in orderto decrease the administration of other treatments or growth factors.

In some embodiments, the 15-PGDH inhibitor can be administered to asubject to enhance recovery of neutrophils following bone marrowtransplantation, following umbilical cord blood transplantation,following transplantation with hematopoietic stem cells, followingconventional chemotherapy, following radiation treatment, and inindividuals with neutropenias from diseases that include but are notlimited to aplastic anemia, myelodysplasia, myelofibrosis, neutropeniasfrom other bone marrow diseases, drug induced neutropenia, immuneneutropenias, idiopathic neutropenia, and following infections withviruses that include, but are not limited to, HIV, CMV, and parvovirus.

In other embodiments, the 15-PGDH inhibitor can be administered to asubject to enhance recovery of platelets following bone marrowtransplantation, following umbilical cord blood transplantation,following transplantation with hematopoietic stem cells, followingconventional chemotherapy, following radiation treatment, and inindividuals with neutropenias from diseases that include but are notlimited to aplastic anemia, myelodysplasia, myelofibrosis,thrombocytopenias from other bone marrow diseases, drug inducedthrombocytopenia, immune thrombocytopenia, idiopathic thrombocytopenicpurpura, idiopathic thrombocytopenia, and following infections withviruses that include, but are not limited to, HIV, CMV, and parvovirus.

In still other embodiments, the 15-PGDH inhibitor can be administered toa subject to enhance recovery of hemoglobin following bone marrowtransplantation, following umbilical cord blood transplantation,following transplantation with hematopoietic stem cells, followingconventional chemotherapy, following radiation treatment, and inindividuals with anemias from diseases that include but are not limitedto aplastic anemia, myelodysplasia, myelofibrosis, anemia from otherbone marrow diseases, drug induced anemia, immune mediated anemias,anemia of chronic disease, idiopathic anemia, and following infectionswith viruses that include, but are not limited to, HIV, CMV, andparvovirus.

In some embodiments, the 15-PGDH inhibitor can be administered to asubject to enhance numbers of bone marrow stem cell numbers followingbone marrow transplantation, following umbilical cord bloodtransplantation, following transplantation with hematopoietic stemcells, following conventional chemotherapy, following radiationtreatment, in individuals with other bone marrow diseases, inindividuals with cytopenias following viral infections, and inindividuals with cytopenias.

In other embodiments, the 15-PGDH inhibitor can be administered to asubject to enhance response to cytokines and/or cell mobilization agentsadministered to individuals with cytopenias that include but are notlimited to neutropenia, thrombocytopenia, lymphocytopenia, and anemia.Cytokines whose responses may be enhanced by 15-PGDH inhibitors include,but are not limited to: G-CSF, GM-CSF, EPO, IL-3, IL-6, TPO, SCF, andTPO-RA (thrombopoietin receptor agonist).

In further embodiments, the 15-PGDH inhibitor can be administered to asubject or to a tissue graft of a subject to mitigate graft rejection,to enhance graft engraftment, to enhance graft engraftment followingtreatment of the subject or the marrow of the subject with radiationtherapy, chemotherapy, or immunosuppressive therapy, to conferresistance to toxic or lethal effects of exposure to radiation, conferresistance to the toxic effect of Cytoxan, the toxic effect offludarabine, the toxic effect of chemotherapy, or the toxic effect ofimmunosuppressive therapy, to decrease infection, and/or to decreasepulmonary toxicity from radiation.

In other embodiments, the 15-PGDH inhibitor can be administered to arecipient of a tissue stem cell transplant, including but not limited toa transplant with hematopoietic stem cells, neural stem stems,mesenchymal stem cells, or stem cells for other tissues, so as toaccelerate tissue regeneration and repair following the transplant.

In some embodiments, the administration of a 15-PGDH inhibitor alone orin combination with a cytokine, hematopoietic cytokine, and/or cellmobilization agent to a subject in need thereof, cells of the subject,and/or tissue of the subject can be used for the purpose of increasingneutrophils in the subject. For example, the hematopoietic cytokine orcell mobilization agent can include at least one of G-CSF or Plerixafor.

In other embodiments, the administration of a 15-PGDH inhibitor alone orin combination with a cytokine, hematopoietic cytokine, and/or cellmobilization agent to a subject in need thereof, cells of the subject,and/or tissue of the subject can be used for the purpose of increasingnumbers of and/or of mobilizing peripheral blood hematopoietic stemcells in the subject. For example, the hematopoietic cytokine or cellmobilization agent can include at least one of G-CSF or Plerixafor.

In still other embodiments, the administration of a 15-PGDH inhibitoralone or in combination with a cytokine, hematopoietic cytokine, and/orcell mobilization agent to a subject in need thereof, cells of thesubject, and/or tissue of the subject can be used for the purpose ofincreasing numbers of hematopoietic stem cells in blood or bone marrowof the subject. For example, the hematopoietic cytokine or cellmobilization agent can include at least one of G-CSF or Plerixafor.

15-PGDH inhibitors can be identified using assays in which putativeinhibitor compounds are applied to cells expressing 15-PGDH and then thefunctional effects on 15-PGDH activity are determined. Samples or assayscomprising 15-PGDH that are treated with a potential inhibitor arecompared to control samples without the inhibitor to examine the extentof effect. Control samples (untreated with modulators) are assigned arelative 15-PGDH activity value of 100%. Inhibition of 15-PGDH isachieved when the 15-PGDH activity value relative to the control isabout 80%, optionally 50% or 25%, 10%, 5% or 1%.

Agents tested as inhibitors of SCD (e.g., 15-PGDH) can be any smallchemical molecule or compound. Typically, test compounds will be smallchemical molecules, natural products, or peptides. The assays aredesigned to screen large chemical libraries by automating the assaysteps and providing compounds from any convenient source to assays,which are typically run in parallel (e.g., in microtiter formats onmicrotiter plates in robotic assays).

In some embodiments, the 15-PGDH inhibitor can include a compound havingthe following formula (I):

wherein n is 0-2;

Y¹, Y², and R¹ are the same or different and are each selected from thegroup consisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O) (C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, arlyl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinY¹ and Y² may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ arethe same or different and are each selected from the group consisting ofH, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

X¹ and X² are independently N or C, and wherein when X¹ and/or X² are N,Y¹ and/or Y², respectively, are absent;

Z¹ is O, S, CR^(a)R^(b) or NR^(a), wherein R^(a) and R^(b) areindependently H or a C₁₋₈ alkyl, which is linear, branched, or cyclic,and which is unsubstituted or substituted;

and pharmaceutically acceptable salts thereof.

Examples of 15-PGDH inhibitors having formulas (I) include the followingcompounds:

and pharmaceutically acceptable salts thereof.

In other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (II):

wherein n is 0-2

X⁴, X⁵, X⁶, and X⁷ are independently N or CR^(c);

R¹, R⁶, R⁷, and R^(c) are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O)(C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinR⁶ and R⁷ may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ arethe same or different and are each selected from the group consisting ofH, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

Z¹ is O, S, CR^(a)R^(b) or NR^(a), wherein R^(a) and R^(b) areindependently H or a C₁₋₈ alkyl, which is linear, branched, or cyclic,and which is unsubstituted or substituted;

and pharmaceutically acceptable salts thereof.

Examples of 15-PGDH inhibitors having formulas (II) include thefollowing compounds:

and pharmaceutically acceptable salts thereof.

In yet other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (III) or (IV):

wherein n is 0-2

X⁶ is independently is N or CR^(c);

R¹, R⁶, R⁷, and R^(c) are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O)(C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinR⁶ and R⁷ may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ arethe same or different and are each selected from the group consisting ofH, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

Z¹ is O, S, CR^(a)R^(b) or NR^(a), wherein R^(a) and R^(b) areindependently H or a C₁₋₈ alkyl, which is linear, branched, or cyclic,and which is unsubstituted or substituted;

and pharmaceutically acceptable salts thereof.

In some embodiments, R¹ is selected from the group consisting ofbranched or linear alkyl including —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5).

In other embodiments, R⁶ and R⁷ can each independently be one of thefollowing:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, and R⁷⁴ are the sameor different and are independently selected from the group consisting ofhydrogen, substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heterocycloalkenyl containing from 5-6 ringatoms, (wherein from 1-3 of the ring atoms is independently selectedfrom N, NH, N(C₁-C₆ alkyl), NC(O) (C₁-C₆ alkyl), O, and S), heteroarylor heterocyclyl containing from 5-14 ring atoms, (wherein from 1-6 ofthe ring atoms is independently selected from N, NH, N(C₁-C₃ alkyl), O,and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, silyl, hydroxyl,sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀aryloxy, acyl (including C₂-C₂₄ alkylcarbonyl (—CO-alkyl) and C₆-C₂₀arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C₂-C₂₄ alkoxycarbonyl(—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl (—(CO)—O-aryl), C₂-C₂₄alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀ arylcarbonato (—O—(CO)—O-aryl),carboxy (—COOH), carboxylato (—COO⁻), carbamoyl (—(CO)—NH₂), C₁-C₂₄alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)), arylcarbamoyl (—(CO)—NH-aryl),thiocarbamoyl (—(CS)—NH₂), carbamido (—NH—(CO)—NH₂), cyano(-CN),isocyano (—N⁺C⁻), cyanato (—O—CN), isocyanato (—O—N⁺═C⁻), isothiocyanato(—S—CN), azido (—N═N⁺═N⁻), formyl (—(CO)—H), thioformyl (—(CS)—H), amino(—NH₂), C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido(—NH—(CO)-alkyl), C₆-C₂₀ arylamido (—NH—(CO)-aryl), sulfanamido(—SO₂N(R)₂ where R is independently H, alkyl, aryl or heteroaryl), imino(—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen,alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), whereR=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO₂), nitroso (—NO),sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄ alkylsulfanyl (—S-alkyl;also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed“arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl), C₅-C₂₀ arylsulfinyl(—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl), C₅-C₂₀ arylsulfonyl(—SO₂-aryl), sulfonamide (—SO₂—NH₂, —SO₂NY₂ (wherein Y is independentlyH, aryl or alkyl), phosphono (—P(O)(OH)₂), phosphonato (—P(O)(O)₂),phosphinato (—P(O)(O⁻)), phospho (—PO₂), phosphino (—PH₂), polyalkylethers (—[(CH₂)_(n)O]_(m)), phosphates, phosphate esters [—OP(O)(OR)₂where R═H, methyl or other alkyl], groups incorporating amino acids orother moieties expected to bear positive or negative charge atphysiological pH, and combinations thereof, and pharmaceuticallyacceptable salts thereof.

In still other embodiments, R⁶ and R⁷ can independently be a group thatimproves aqueous solubility, for example, a phosphate ester (—OPO₃H₂), aphenyl ring linked to a phosphate ester (—OPO₃H₂), a phenyl ringsubstituted with one or more methoxyethoxy groups, or a morpholine, oran aryl or heteroaryl ring substituted with such a group.

Examples of 15-PGDH inhibitors having formulas (III) or (IV) include thefollowing compounds:

and pharmaceutically acceptable salts thereof.

In other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (V):

wherein n is 0-2

X⁶ is independently is N or CR^(c)

R¹, R⁶, R⁷, and R^(c) are each independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O)(C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinR⁶ and R⁷ may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl;

U¹ is N, C—R², or C—NR³R⁴, wherein R² is selected from the groupconsisting of a H, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1,2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X, CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, O(CO)R′,COOR′ (wherein R′ is H or a lower alkyl group), and wherein R¹ and R²may be linked to form a cyclic or polycyclic ring, wherein R³ and R⁴ arethe same or different and are each selected from the group consisting ofH, a lower alkyl group, O, (CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃,CH₂—CH₂X, CH₂—CH₂—CH₂X, (wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′,(C═O)N(R′)₂, COOR′ (wherein R′ is H or a lower alkyl group), and R³ orR⁴ may be absent;

and pharmaceutically acceptable salts thereof.

In some embodiments, R¹ is selected from the group consisting ofbranched or linear alkyl including —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5).

In other embodiments, R⁶ and R⁷ can each independently be one of thefollowing:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, and R⁷⁴, are the sameor different and are independently selected from the group consisting ofhydrogen, substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heterocycloalkenyl containing from 5-6 ringatoms, (wherein from 1-3 of the ring atoms is independently selectedfrom N, NH, N(C₁-C₆ alkyl), NC(O)(C₁-C₆ alkyl), O, and S), heteroaryl orheterocyclyl containing from 5-14 ring atoms, (wherein from 1-6 of thering atoms is independently selected from N, NH, N(C₁-C₃ alkyl), O, andS), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, silyl, hydroxyl, sulfhydryl,C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy,acyl (including C₂-C₂₄ alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl(—CO-aryl)), acyloxy (—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl),C₆-C₂₀ aryloxycarbonyl (—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato(—O—(CO)—O-alkyl), C₆-C₂₀ arylcarbonato (—O—(CO)—O-aryl), carboxy(—COOH), carboxylato (—COO⁻), carbamoyl (—(CO)—NH₂), C₁-C₂₄alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)), arylcarbamoyl (—(CO)—NH-aryl),thiocarbamoyl (—(CS)—NH₂), carbamido (—NH—(CO)—NH₂), cyano(-CN),isocyano (—N⁺C⁻), cyanato (—O—CN), isocyanato (—O—N⁺═C⁻), isothiocyanato(—S—CN), azido (—N═N⁺═N⁻), formyl (—(CO)—H), thioformyl (—(CS)—H), amino(—NH₂), C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido(—NH—(CO)-alkyl), C₆-C₂₀ arylamido (—NH—(CO)-aryl), sulfanamido(—SO₂N(R)₂ where R is independently H, alkyl, aryl or heteroaryl), imino(—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl), where R=hydrogen,alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (—CR═N(aryl), whereR=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO₂), nitroso (—NO),sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄ alkylsulfanyl (—S-alkyl;also termed “alkylthio”), arylsulfanyl (—S-aryl; also termed“arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl), C₅-C₂₀ arylsulfinyl(—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl), C₅-C₂₀ arylsulfonyl(—SO₂-aryl), sulfonamide (—SO₂—NH₂, —SO₂NY₂ (wherein Y is independentlyH, aryl or alkyl), phosphono (—P(O)(OH)₂), phosphonato (—P(O)(O⁻)₂),phosphinato (—P(O)(O⁻)), phospho (—PO₂), phosphino (—PH₂), polyalkylethers (—[(CH₂)_(n)O]_(m)), phosphates, phosphate esters [—OP(O)(OR)₂where R═H, methyl or other alkyl], groups incorporating amino acids orother moieties expected to bear positive or negative charge atphysiological pH, and combinations thereof, and pharmaceuticallyacceptable salts thereof.

In still other embodiments, R⁶ and R⁷ can independently be a group thatimproves aqueous solubility, for example, a phosphate ester (—OPO₃H₂), aphenyl ring linked to a phosphate ester (—OPO₃H₂), a phenyl ringsubstituted with one or more methoxyethoxy groups, or a morpholine, oran aryl or heteroaryl ring substituted with such a group.

In other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (VI):

wherein n=0-2;

X⁶ is N or CR^(c);

R¹ is selected from the group consisting of branched or linear alkylincluding —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5).

R⁵ is selected from the group consisting of H, Cl, F, NH₂, and N(R⁷⁶)₂;

R⁶ and R⁷ can each independently be one of the following:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, R⁷⁴, R⁷⁶, and R^(c)are the same or different and are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heterocycloalkenylcontaining from 5-6 ring atoms, (wherein from 1-3 of the ring atoms isindependently selected from N, NH, N(C₁-C₆ alkyl), NC(O) (C₁-C₆ alkyl),O, and S), heteroaryl or heterocyclyl containing from 5-14 ring atoms,(wherein from 1-6 of the ring atoms is independently selected from N,NH, N(C₁-C₃ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,silyl, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄ alkylcarbonyl(—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl),C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), sulfanamido (—SO₂N(R)₂ where R is independently H,alkyl, aryl or heteroaryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄alkyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino(—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.),arylimino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.),nitro (—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻),C₁-C₂₄ alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH₂, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkyl ethers (—[(CH₂)_(n)O]_(m)), phosphates,phosphate esters [—OP(O)(OR)₂ where R═H, methyl or other alkyl], groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, and combinations thereof, andpharmaceutically acceptable salts thereof.

In other embodiments, the 15-PGDH inhibitor can include a compoundhaving the following formula (VII):

wherein n=0-2;

X⁶ is N or CR^(c);

R¹ is selected from the group consisting of branched or linear alkylincluding —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5).

R⁵ is selected from the group consisting of H, Cl, F, NH₂, and N(R⁷⁶)₂;

R⁷ can each independently be one of the following:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, R⁷⁴, R⁷⁶, and R^(c)are the same or different and are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heterocycloalkenylcontaining from 5-6 ring atoms, (wherein from 1-3 of the ring atoms isindependently selected from N, NH, N(C₁-C₆ alkyl), NC(O)(C₁-C₆ alkyl),O, and S), heteroaryl or heterocyclyl containing from 5-14 ring atoms,(wherein from 1-6 of the ring atoms is independently selected from N,NH, N(C₁-C₃ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,silyl, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄ alkylcarbonyl(—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl),C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂), carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formylthioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino, C₅-C₂₀ arylamino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), sulfanamido (—SO₂N(R)₂ where R is independently H,alkyl, aryl or heteroaryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄alkyl, C₅-C₂₀ aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino(—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.),arylimino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.),nitro (—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻),C₁-C₂₄ alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH₂, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkyl ethers (—[(CH₂)_(n)O]_(m)), phosphates,phosphate esters [—OP(O)(OR)₂ where R═H, methyl or other alkyl], groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, and combinations thereof, andpharmaceutically acceptable salts thereof.

Examples of compounds having formulas (V), (VI), or (VII) are selectedfrom the group consisting of:

and pharmaceutically acceptable salts thereof.

In certain embodiments, the 15-PGDH inhibitor having formula (I), (II),(III), (IV), (V), (VI), and (VII) can be selected that can ia) at 2.5 μMconcentration, stimulate a Vaco503 reporter cell line expressing a15-PGDH luciferase fusion construct to a luciferase output level ofgreater than 70 (using a scale on which a value of 100 indicates adoubling of reporter output over baseline); iia) at 2.5 μM concentrationstimulate a V9m reporter cell line expressing a 15-PGDH luciferasefusion construct to a luciferase output level of greater than 75; iiia)at 7.5 μM concentration stimulate a LS174T reporter cell line expressinga 15-PGDH luciferase fusion construct to a luciferase output level ofgreater than 70; and iva) at 7.5 μM concentration, does not activate anegative control V9m cell line expressing TK-renilla luciferase reporterto a level greater than 20; and va) inhibits the enzymatic activity ofrecombinant 15-PGDH protein at an IC₅₀ of less than 1 μM.

In other embodiments, the 15-PGDH inhibitor can ib) at 2.5 μMconcentration, stimulate a Vaco503 reporter cell line expressing a15-PGDH luciferase fusion construct to increase luciferase output; iib)at 2.5 μM concentration stimulate a V9m reporter cell line expressing a15-PGDH luciferase fusion construct to increase luciferase output; iiib)at 7.5 μM concentration stimulate a LS174T reporter cell line expressinga 15-PGDH luciferase fusion construct to increase luciferase output;ivb) at 7.5 μM concentration, does not activate a negative control V9mcell line expressing TK-renilla luciferase reporter to a luciferaselevel greater than 20% above background; and vb) inhibits the enzymaticactivity of recombinant 15-PGDH protein at an IC₅₀ of less than 1 μM.

In other embodiments, the 15-PGDH inhibitor can inhibit the enzymaticactivity of recombinant 15-PGDH at an IC₅₀ of less than 1 μM, orpreferably at an IC₅₀ of less than 250 nM, or more preferably at an IC₅₀of less than 50 nM, or more preferably at an IC₅₀ of less than 10 nM, ormore preferably at an IC₅₀ of less than 5 nM at a recombinant 15-PGDHconcentration of about 5 nM to about 10 nM.

In other embodiments, the 15-PGDH inhibitor can increase the cellularlevels of PGE-2 following stimulation of an A459 cell with anappropriate agent, for example IL1-beta.

In some embodiments, a15-PGDH inhibitor can include a compound havingthe following formula (VIII):

wherein n is 0-2;

R¹, R⁶, and R⁷ are the same or different and are each selected from thegroup consisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms (wherein from 1-3 ofthe ring atoms is independently selected from N, NH, N(C₁-C₆ alkyl),NC(O) (C₁-C₆ alkyl), O, and S), C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo,—Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl (including C₂-C₂₄alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl (—CO-aryl)), acyloxy(—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl), C₆-C₂₀ aryloxycarbonyl(—(CO)—O-aryl), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀arylcarbonato (—O—(CO)—O-aryl), carboxy (—COOH), carboxylato (—COO⁻),carbamoyl (—(CO)—NH₂), C₁-C₂₄ alkyl-carbamoyl (—(CO)—NH(C₁-C₂₄ alkyl)),arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl (—(CS)—NH₂) carbamido(—NH—(CO)—NH₂), cyano(-CN), isocyano (—N⁺C⁻), cyanato (—O—CN),isocyanato (—O—N⁺═C⁻), isothiocyanato (—S—CN), azido (—N═N⁺═N⁻), formyl(—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₆-C₂₀ arylamido(—NH—(CO)-aryl), imino (—CR═NH where R is hydrogen, C₁-C₂₄ alkyl, C₅-C₂₀aryl, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, etc.), alkylimino (—CR═N(alkyl),where R=hydrogen, alkyl, aryl, alkaryl, aralkyl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O⁻), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), sulfonamide (—SO₂—NH2, —SO₂NY₂ (whereinY is independently H, aryl or alkyl), phosphono (—P(O)(OH)₂),phosphonato (—P(O)(O⁻)₂), phosphinato (—P(O)(O⁻)), phospho (—PO₂),phosphino (—PH₂), polyalkylethers, phosphates, phosphate esters, groupsincorporating amino acids or other moieties expected to bear positive ornegative charge at physiological pH, combinations thereof, and whereinR⁶ and R⁷ may be linked to form a cyclic or polycyclic ring, wherein thering is a substituted or unsubstituted aryl, a substituted orunsubstituted heteroaryl, a substituted or unsubstituted cycloalkyl, anda substituted or unsubstituted heterocyclyl; and pharmaceuticallyacceptable salts thereof.

15-PGDH inhibitors having formula (VIII) can be synthesized as shown:

Any reaction solvent can be used in the above preparation process aslong as it is not involved in the reaction. For example, the reactionsolvent includes ethers such as diethyl ether, tetrahydrofuran anddioxane; halogenized hydrocarbons, such as dichloromethane andchloroform; amines such as pyridine, piperidine and triethylamine;alkylketones, such as acetone, methylethylketone and methylisobutyl;alcohols, such as methanol, ethanol and propanol; non-protonic polarsolvent, such as N,N-dimethylformamide, N,N-dimethylacetamide,acetonitrile, dimethylsulfoxide and hexamethyl phosphoric acid triamide.Among non-reactive organic solvents that are ordinarily used in theorganic synthesis, preferable solvents are those from which watergenerated in the reaction can be removed by a Dean-Stark trap. Theexamples of such solvents include, but are not limited to benzene,toluene, xylene and the like. The reaction product thus obtained may beisolated and purified by condensation, extraction and the like, which isordinarily conducted in the field of the organic synthesis, if desired,by silica gel column chromatography. The individual enantiomers of PGDHinhibitors having the formula III can be separated by a preparative HPLCusing chromatography columns containing chiral stationary phases.

Further, embodiments of this application include any modifications forthe preparation method of the 15-PGDH inhibitors described above. Inthis connection, any intermediate product obtainable from any step ofthe preparation method can be used as a starting material in the othersteps. Such starting material can be formed in situ under certainreaction conditions. Reaction reagents can also be used in the form oftheir salts or optical isomers.

Depending on the kinds of the substituents to be used in the preparationof the 15-PGDH inhibitors, and the intermediate product and thepreparation method selected, novel 15-PGDH inhibitors can be in the formof any possible isomers such as substantially pure geometrical (cis ortrans) isomers, optical isomers (enantiomers) and racemates.

In some embodiments, a 15-PGDH inhibitor having formula (VIII) caninclude a compound with the following formula (IX):

and pharmaceutically acceptable salts thereof.

Advantageously, the 15-PDGH inhibitor having formula (IX) was found to:i) inhibit recombinant 15-PGDH at 1 nM concentration; ii) inhibit15-PGDH in cell lines at 100 nM concentration, iii) increase PGE₂production by cell lines; iv) is chemically stable in aqueous solutionsover broad pH range; v) is chemically stable when incubated withhepatocyte extracts, vi) is chemically stable when incubated withhepatocyte cell lines; vii) shows 253 minutes plasma half-life wheninjected IP into mice; and viii) shows no immediate toxicity over 24hours when injected IP into mice at 0.6 μmole/per mouse and at 1.2μmole/per mouse and also no toxicity when injected IP into mice at 0.3μmole/per mouse twice daily for 21 days.

In other embodiments, a 15-PGDH inhibitor having formula (IX) caninclude a compound with the following formula (IXa):

and pharmaceutically acceptable salts thereof.

In still other embodiments, a 15-PGDH inhibitor having formula (IX) caninclude a compound with the following formula (IXb):

and pharmaceutically acceptable salts thereof.

In other embodiments, the 15-PDHG inhibitor can comprise a (+) or (−)optical isomer of a 15-PGDH inhibitor having formula (IX). In stillother embodiments, the 15-PDHG inhibitor can comprise a mixture at leastone of a (+) or (−) optical isomer of a 15-PGDH inhibitor having formula(IX). For example, the 15-PGDH inhibitor can comprise a mixture of: lessthan about 50% by weight of the (−) optical isomer of a 15-PGDHinhibitor having formula (IX) and greater than about 50% by weight ofthe (+) optical isomer of a 15-PGDH inhibitor having formula (IX), lessthan about 25% by weight of the (−) optical isomer of a 15-PGDHinhibitor having formula (IX) and greater than about 75% by weight ofthe (+) optical isomer of a 15-PGDH inhibitor having formula (IX), lessthan about 10% by weight of the (−) optical isomer of a 15-PGDHinhibitor having formula (IX) and greater than about 90% by weight ofthe (+) optical isomer of a 15-PGDH inhibitor having formula (IX), lessthan about 1% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (IX) and greater than about 99% by weight of the (+)optical isomer of a 15-PGDH inhibitor having formula (IX), greater thanabout 50% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (IX) and less than about 50% by weight of the (+) opticalisomer of a 15-PGDH inhibitor having formula (IX), greater than about75% by weight of the (−) optical isomer of a 15-PGDH inhibitor havingformula (IX) and less than about 25% by weight of the (+) optical isomerof a 15-PGDH inhibitor having formula (IX), greater than about 90% byweight of the (−) optical isomer of a 15-PGDH inhibitor having formula(IX) and less than about 10% by weight of the (+) optical isomer of a15-PGDH inhibitor having formula (IX), or greater than about 99% byweight of the (−) optical isomer of a 15-PGDH inhibitor having formula(IX) and less than about 1% by weight of the (+) optical isomer of a15-PGDH inhibitor having formula (IX).

In a still further embodiment, the 15-PDGH inhibitor can consistessentially of or consist of the (+) optical isomer of a 15-PGDHinhibitor having formula (IX). In yet another embodiment, the PDGHinhibitor can consist essentially of or consist of the (−) opticalisomer of a 15-PGDH inhibitor having formula (IX).

In other embodiments, a 15-PGDH inhibitor having formula (VIII) caninclude a compound with the following formula (X):

and pharmaceutically acceptable salts thereof.

Advantageously, the 15-PDGH inhibitor having formula (X) was found to:i) inhibit recombinant 15-PGDH at 3 nM concentration; ii) increase PGE₂production by cell lines at 20 nM; iii) is chemically stable in aqueoussolutions over broad pH range; iv) is chemically stable when incubatedwith mouse, rat and human liver extracts, v) shows 33 minutes plasmahalf-life when injected IP into mice; viii) shows no immediate toxicityover 24 hours when injected IP into mice at 50 mg/kg body weight, andix) is soluble in water (pH=3) at 1 mg/mL.

In other embodiments, a 15-PGDH inhibitor having formula (X) can includea compound with the following formula (Xa):

and pharmaceutically acceptable salts thereof.

In still other embodiments, a 15-PGDH inhibitor having formula (X) caninclude a compound with the following formula (Xb):

and pharmaceutically acceptable salts thereof.

In other embodiments, the 15-PDHG inhibitor can comprise a (+) or (−)optical isomer of a 15-PGDH inhibitor having formula (X). In still otherembodiments, the 15-PDHG inhibitor can comprise a mixture at least oneof a (+) or (−) optical isomer of a 15-PGDH inhibitor having formula(X). For example, the 15-PGDH inhibitor can comprise a mixture of: lessthan about 50% by weight of the (−) optical isomer of a 15-PGDHinhibitor having formula (X) and greater than about 50% by weight of the(+) optical isomer of a 15-PGDH inhibitor having formula (X), less thanabout 25% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (X) and greater than about 75% by weight of the (+)optical isomer of a 15-PGDH inhibitor having formula (X), less thanabout 10% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (X) and greater than about 90% by weight of the (+)optical isomer of a 15-PGDH inhibitor having formula (X), less thanabout 1% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (X) and greater than about 99% by weight of the (+)optical isomer of a 15-PGDH inhibitor having formula (X), greater thanabout 50% by weight of the (−) optical isomer of a 15-PGDH inhibitorhaving formula (X) and less than about 50% by weight of the (+) opticalisomer of a 15-PGDH inhibitor having formula (X), greater than about 75%by weight of the (−) optical isomer of a 15-PGDH inhibitor havingformula (X) and less than about 25% by weight of the (+) optical isomerof a 15-PGDH inhibitor having formula (X), greater than about 90% byweight of the (−) optical isomer of a 15-PGDH inhibitor having formula(X) and less than about 10% by weight of the (+) optical isomer of a15-PGDH inhibitor having formula (X), or greater than about 99% byweight of the (−) optical isomer of a 15-PGDH inhibitor having formula(X) and less than about 1% by weight of the (+) optical isomer of a15-PGDH inhibitor having formula (X).

In a still further embodiment, the 15-PDGH inhibitor can consistessentially of or consist of the (+) optical isomer of a 15-PGDHinhibitor having formula (X). In yet another embodiment, the PDGHinhibitor can consist essentially of or consist of the (−) opticalisomer of a 15-PGDH inhibitor having formula (X).

It will be appreciated that the other 15-PGDH inhibitors can be used inthe methods described herein. These other 15-PGDH inhibitors can includeknown 15-PGDH inhibitors including, for example, tetrazole compounds offormulas (I) and (II), 2-alkylideneaminooxyacetamide compounds offormula (I), heterocyclic compounds of formulas (VI) and (VII), andpyrazole compounds of formula (III) described in U.S. Patent ApplicationPublication No. 2006/0034786 and U.S. Pat. No. 7,705,041;benzylidene-1,3-thiazolidine compounds of formula (I) described in U.S.Patent Application Publication No. 2007/0071699;phenylfurylmethylthiazolidine-2,4-dione andphenylthienylmethylthiazolidine-2,4-dione compounds described in U.S.Patent Application Publication No. 2007/0078175; thiazolidenedionederivatives described in U.S. Patent Application Publication No.2011/0269954; phenylfuran, phenylthiophene, or phenylpyrrazole compoundsdescribed in U.S. Pat. No. 7,294,641, 5-(3,5-disubstitutedphenylazo)-2-hydroxybenzene-acetic acids and salts and lactonesdescribed in U.S. Pat. No. 4,725,676, and azo compounds described inU.S. Pat. No. 4,889,846.

The 15-PGDH inhibitors described herein can be provided in apharmaceutical composition or cosmetic composition depending on thepathological or cosmetic condition or disorder being treated. Apharmaceutical composition containing the 15-PGDH inhibitors describedherein as an active ingredient may be manufactured by mixing thederivative with a pharmaceutically acceptable carrier(s) or anexcipient(s) or diluting the 15-PGDH inhibitors with a diluent inaccordance with conventional methods. The pharmaceutical composition mayfurther contain fillers, anti-cohesives, lubricants, wetting agents,flavoring agents, emulsifying agents, preservatives and the like. Thepharmaceutical composition may be formulated into a suitable formulationin accordance with the methods known to those skilled in the art so thatit can provide an immediate, controlled or sustained release of the15-PGDH inhibitors after being administered into a mammal.

In some embodiments, the pharmaceutical composition may be formulatedinto a parenteral or oral dosage form. The solid dosage form for oraladministration may be manufactured by adding excipient, if necessary,together with binder, disintegrants, lubricants, coloring agents, and/orflavoring agents, to the 15-PGDH inhibitors and shaping the resultingmixture into the form of tablets, sugar-coated pills, granules, powderor capsules. The additives that can be added in the composition may beordinary ones in the art. For example, examples of the excipient includelactose, sucrose, sodium chloride, glucose, starch, calcium carbonate,kaolin, microcrystalline cellulose, silicate and the like. Exemplarybinders include water, ethanol, propanol, sweet syrup, sucrose solution,starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose,shellac, calcium phosphonate and polypyrrolidone. Examples of thedisintegrant include dry starch, sodium arginate, agar powder, sodiumbicarbonate, calcium carbonate, sodium lauryl sulfate, stearicmonoglyceride and lactose. Further, purified talc, stearates, sodiumborate, and polyethylene glycol may be used as a lubricant; and sucrose,bitter orange peel, citric acid, tartaric acid, may be used as aflavoring agent. In some embodiments, the pharmaceutical composition canbe made into aerosol formulations (e.g., they can be nebulized) to beadministered via inhalation.

The 15-PGDH inhibitors described herein may be combined with flavoringagents, buffers, stabilizing agents, and the like and incorporated intooral liquid dosage forms such as solutions, syrups or elixirs inaccordance with conventional methods. One example of the buffers may besodium citrate. Examples of the stabilizing agents include tragacanth,acacia and gelatin.

In some embodiments, the 15-PGDH inhibitors described herein may beincorporated into an injection dosage form, for example, for asubcutaneous, intramuscular or intravenous route by adding thereto pHadjusters, buffers, stabilizing agents, relaxants, topical anesthetics.Examples of the pH adjusters and the buffers include sodium citrate,sodium acetate and sodium phosphate. Examples of the stabilizing agentsinclude sodium pyrosulfite, EDTA, thioglycolic acid and thiolactic acid.The topical anesthetics may be procaine HCl, lidocaine HCl and the like.The relaxants may be sodium chloride, glucose and the like.

In other embodiments, the 15-PGDH inhibitors described herein may beincorporated into suppositories in accordance with conventional methodsby adding thereto pharmaceutically acceptable carriers that are known inthe art, for example, polyethylene glycol, lanolin, cacao butter orfatty acid triglycerides, if necessary, together with surfactants suchas Tween.

The pharmaceutical composition may be formulated into various dosageforms as discussed above and then administered through various routesincluding an oral, inhalational, transdermal, subcutaneous, intravenousor intramuscular route. The dosage can be a pharmaceutically ortherapeutically effective amount.

Therapeutically effective dosage amounts of the 15-PGDH inhibitor may bepresent in varying amounts in various embodiments. For example, in someembodiments, a therapeutically effective amount of the 15-PGDH inhibitormay be an amount ranging from about 10-1000 mg (e.g., about 20 mg-1,000mg, 30 mg-1,000 mg, 40 mg-1,000 mg, 50 mg-1,000 mg, 60 mg-1,000 mg, 70mg-1,000 mg, 80 mg-1,000 mg, 90 mg-1,000 mg, about 10-900 mg, 10-800 mg,10-700 mg, 10-600 mg, 10-500 mg, 100-1000 mg, 100-900 mg, 100-800 mg,100-700 mg, 100-600 mg, 100-500 mg, 100-400 mg, 100-300 mg, 200-1000 mg,200-900 mg, 200-800 mg, 200-700 mg, 200-600 mg, 200-500 mg, 200-400 mg,300-1000 mg, 300-900 mg, 300-800 mg, 300-700 mg, 300-600 mg, 300-500 mg,400 mg-1,000 mg, 500 mg-1,000 mg, 100 mg-900 mg, 200 mg-800 mg, 300mg-700 mg, 400 mg-700 mg, and 500 mg-600 mg). In some embodiments, the15-PGDH inhibitor is present in an amount of or greater than about 10mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg. In someembodiments, the 15-PGDH inhibitor is present in an amount of or lessthan about 1000 mg, 950 mg, 900 mg, 850 mg, 800 mg, 750 mg, 700 mg, 650mg, 600 mg, 550 mg, 500 mg, 450 mg, 400 mg, 350 mg, 300 mg, 250 mg, 200mg, 150 mg, or 100 mg.

In other embodiments, a therapeutically effective dosage amount may be,for example, about 0.001 mg/kg weight to 500 mg/kg weight, e.g., fromabout 0.001 mg/kg weight to 400 mg/kg weight, from about 0.001 mg/kgweight to 300 mg/kg weight, from about 0.001 mg/kg weight to 200 mg/kgweight, from about 0.001 mg/kg weight to 100 mg/kg weight, from about0.001 mg/kg weight to 90 mg/kg weight, from about 0.001 mg/kg weight to80 mg/kg weight, from about 0.001 mg/kg weight to 70 mg/kg weight, fromabout 0.001 mg/kg weight to 60 mg/kg weight, from about 0.001 mg/kgweight to 50 mg/kg weight, from about 0.001 mg/kg weight to 40 mg/kgweight, from about 0.001 mg/kg weight to 30 mg/kg weight, from about0.001 mg/kg weight to 25 mg/kg weight, from about 0.001 mg/kg weight to20 mg/kg weight, from about 0.001 mg/kg weight to 15 mg/kg weight, fromabout 0.001 mg/kg weight to 10 mg/kg weight.

In still other embodiments, a therapeutically effective dosage amountmay be, for example, about 0.0001 mg/kg weight to 0.1 mg/kg weight, e.g.from about 0.0001 mg/kg weight to 0.09 mg/kg weight, from about 0.0001mg/kg weight to 0.08 mg/kg weight, from about 0.0001 mg/kg weight to0.07 mg/kg weight, from about 0.0001 mg/kg weight to 0.06 mg/kg weight,from about 0.0001 mg/kg weight to 0.05 mg/kg weight, from about 0.0001mg/kg weight to about 0.04 mg/kg weight, from about 0.0001 mg/kg weightto 0.03 mg/kg weight, from about 0.0001 mg/kg weight to 0.02 mg/kgweight, from about 0.0001 mg/kg weight to 0.019 mg/kg weight, from about0.0001 mg/kg weight to 0.018 mg/kg weight, from about 0.0001 mg/kgweight to 0.017 mg/kg weight, from about 0.0001 mg/kg weight to 0.016mg/kg weight, from about 0.0001 mg/kg weight to 0.015 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.014 mg/kg weight, from about 0.0001 mg/kgweight to 0.013 mg/kg weight, from about 0.0001 mg/kg weight to 0.012mg/kg weight, from about 0.0001 mg/kg weight to 0.011 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.01 mg/kg weight, from about 0.0001 mg/kgweight to 0.009 mg/kg weight, from about 0.0001 mg/kg weight to 0.008mg/kg weight, from about 0.0001 mg/kg weight to 0.007 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.006 mg/kg weight, from about 0.0001 mg/kgweight to 0.005 mg/kg weight, from about 0.0001 mg/kg weight to 0.004mg/kg weight, from about 0.0001 mg/kg weight to 0.003 mg/kg weight, fromabout 0.0001 mg/kg weight to 0.002 mg/kg weight. In some embodiments,the therapeutically effective dose may be 0.0001 mg/kg weight, 0.0002mg/kg weight, 0.0003 mg/kg weight, 0.0004 mg/kg weight, 0.0005 mg/kgweight, 0.0006 mg/kg weight, 0.0007 mg/kg weight, 0.0008 mg/kg weight,0.0009 mg/kg weight, 0.001 mg/kg weight, 0.002 mg/kg weight, 0.003 mg/kgweight, 0.004 mg/kg weight, 0.005 mg/kg weight, 0.006 mg/kg weight,0.007 mg/kg weight, 0.008 mg/kg weight, 0.009 mg/kg weight, 0.01 mg/kgweight, 0.02 mg/kg weight, 0.03 mg/kg weight, 0.04 mg/kg weight, 0.05mg/kg weight, 0.06 mg/kg weight, 0.07 mg/kg weight, 0.08 mg/kg weight,0.09 mg/kg weight, or 0.1 mg/kg weight. The effective dose for aparticular individual can be varied (e.g., increased or decreased) overtime, depending on the needs of the individual.

In some embodiments, a therapeutically effective dosage may be a dosageof 10 μg/kg/day, 50 μg/kg/day, 100 μg/kg/day, 250 μg/kg/day, 500μg/kg/day, 1000 μg/kg/day or more. In various embodiments, the amount ofthe 15-PGDH inhibitor or pharmaceutical salt thereof is sufficient toprovide a dosage to a patient of between 0.01 μg/kg and 10 μg/kg; 0.1μg/kg and 5 μg/kg; 0.1 μg/kg and 1000 μg/kg; 0.1 μg/kg and 900 μg/kg;0.1 μg/kg and 900 μg/kg; 0.1 μg/kg and 800 μg/kg; 0.1 μg/kg and 700μg/kg; 0.1 μg/kg and 600 μg/kg; 0.1 μg/kg and 500 μg/kg; or 0.1 μg/kgand 400 μg/kg.

Particular doses or amounts to be administered in accordance with thepresent invention may vary, for example, depending on the nature and/orextent of the desired outcome, on particulars of route and/or timing ofadministration, and/or on one or more characteristics (e.g., weight,age, personal history, genetic characteristic, lifestyle parameter,severity of cardiac defect and/or level of risk of cardiac defect, etc.,or combinations thereof). Such doses or amounts can be determined bythose of ordinary skill. In some embodiments, an appropriate dose oramount is determined in accordance with standard clinical techniques.For example, in some embodiments, an appropriate dose or amount is adose or amount sufficient to reduce a disease severity index score by 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100% or more.For example, in some embodiments, an appropriate dose or amount is adose or amount sufficient to reduce a disease severity index score by 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100%.Alternatively or additionally, in some embodiments, an appropriate doseor amount is determined through use of one or more in vitro or in vivoassays to help identify desirable or optimal dosage ranges or amounts tobe administered.

Various embodiments may include differing dosing regimen. In someembodiments, the 15-PGDH inhibitor can be administered via continuousinfusion. In some embodiments, the continuous infusion is intravenous.In other embodiments, the continuous infusion is subcutaneous.Alternatively or additionally, in some embodiments, the 15-PGDHinhibitor can be administered bimonthly, monthly, twice monthly,triweekly, biweekly, weekly, twice weekly, thrice weekly, daily, twicedaily, or on another clinically desirable dosing schedule. The dosingregimen for a single subject need not be at a fixed interval, but can bevaried over time, depending on the needs of the subject.

For topical application, the composition can be administered in the formof aqueous, alcoholic, aqueous-alcoholic or oily solutions orsuspensions, or of a dispersion of the lotion or serum type, ofemulsions that have a liquid or semi-liquid consistency or are pasty,obtained by dispersion of a fatty phase in an aqueous phase (0/W) orvice versa (W/O) or multiple emulsions, of a free or compacted powder tobe used as it is or to be incorporated into a physiologically acceptablemedium, or else of microcapsules or microparticles, or of vesiculardispersions of ionic and/or nonionic type. It may thus be in the form ofa salve, a tincture, milks, a cream, an ointment, a powder, a patch, animpregnated pad, a solution, an emulsion or a vesicular dispersion, alotion, aqueous or anhydrous gels, a spray, a suspension, a shampoo, anaerosol or a foam. It may be anhydrous or aqueous. It may also comprisesolid preparations constituting soaps or cleansing cakes.

Pharmaceutical compositions including the 15-PGDH inhibitor describedherein can additionally contain, for example, at least one compoundchosen from prostaglandins, in particular prostaglandin PGE₁, PGE₂,their salts, their esters, their analogues and their derivatives, inparticular those described in WO 98/33497, WO 95/11003, JP 97-100091, JP96-134242, in particular agonists of the prostaglandin receptors. It mayin particular contain at least one compound such as the agonists (inacid form or in the form of a precursor, in particular in ester form) ofthe prostaglandin F₂α receptor, such as for example latanoprost,fluprostenol, cloprostenol, bimatoprost, unoprostone, the agonists (andtheir precursors, in particular the esters such as travoprost) of theprostaglandin E₂ receptors such as 17-phenyl PGE₂, viprostol, butaprost,misoprostol, sulprostone, 16,16-dimethyl PGE₂, 11-deoxy PGE₁, 1-deoxyPGE₁, the agonists and their precursors, in particular esters, of theprostacycline (IP) receptor such as cicaprost, iloprost,isocarbacycline, beraprost, eprostenol, treprostinil, the agonists andtheir precursors, in particular the esters, of the prostaglandin D₂receptor such as BW245C((4S)-(3-[(3R,S)-3-cyclohexyl-3-isopropyl]-2,5-dioxo)-4-imidazolidinehept-anoicacid), BW246C((4R)-(3-[(3R,S)-3-cyclohexyl-3-isopropyl]-2,5-dioxo)-4-imidazolidinehept-anoicacid), the agonists and their precursors, in particular the esters, ofthe receptor for the thromboxanes A2 (TP) such as I-BOP ([1S-[1a,2a(Z),3b(1E,3S),4a]]-7-[3-[3-hydroxy-4-[4-(iodophenoxy)-1-butenyl]-7-oxabicyclo-[2.2.1]hept-2-yl]-5-heptenoicacid).

Advantageously, the composition can include at least one 15-PGDHinhibitor as defined above and at least one prostaglandin or oneprostaglandin derivative such as for example the prostaglandins ofseries 2 including in particular PGF₂α and PGE₂ in saline form or in theform of precursors, in particular of the esters (example isopropylesters), their derivatives such as 16,16-dimethyl PGE₂, 17-phenyl PGE₂and 16,16-dimethyl PGF_(2α) 17-phenyl PGF_(2α), prostaglandins of series1 such as 11-deoxyprostaglandin E1, 1-deoxyprostaglandin E1 in saline orester form, is their analogues, in particular latanoprost, travoprost,fluprostenol, unoprostone, bimatoprost, cloprostenol, viprostol,butaprost, misoprostol, their salts or their esters.

The invention is further illustrated by the following examples, which isnot intended to limit the scope of the claims.

Example 1

Analysis of Effect of SW033291 on Bone Marrow Function

This Example shows effects of SW033291 on bone marrow function.

FIGS. 1(A-C) show analysis of bone marrow of wild-type mice versus micethat are homozygous genetic knockouts for 15-PGDH (PGDH−/− mice). Totalbone marrow cellularity and percent of Scal+/c-Kit+ cells in lineagenegative (SKL) cells are the same in both sets of mice. However, bonemarrow from 15-PGDH−/− mice shows an approximately 50% increase innumbers of hematopoietic colonies generated when marrow is plated intomethylcelluose. 15-PGDH knockout mice are denoted by label PGDH mice andby label 15-PGDH. WT denotes wild-type mice.

FIGS. 2(A-F) shows assays in which bone marrow is harvested from awild-type mouse, and incubated ex vivo on ice for 2 hours with eitherSW033291 (0.5 μM), or 1 μM PGE2 or 1 μM 16,16-dimethyl PGE2 (dmPGE2).Treated marrow is again then plated into methylcellulose for counting ofhematopoietic colonies. SW033291 treated marrow again shows anapproximately 50% increase in the number of bone marrow derived coloniesgenerated. Under these conditions, a lesser increase is seen in marrowtreated with PGE2, and a slightly greater increase is seen in marrowtreated with dmPGE2.

FIGS. 3(A-C) show a study of C57BL/6J mice treated with IP SW033291administered in a vehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W at adose of 5 mg/kg or 20 mg/kg. Panel A shows mouse bone marrowcellularity, white blood count (wbc), red blood count (rbc) andplatelets counts. Panel B shows percent of Scal+/c-Kit+ cells in lineagenegative (SKL) cells are unchanged in SW033291 treated mice. Panel Cshows that marrow from SW033291 treated mice gives rise to approximately30% increase in numbers of hematopoietic colonies generated when marrowis plated into methylcelluose. Experimental conditions are noted on thefigure.

FIGS. 4(A-B) show analysis of marrow from CD45.2 antigen marked C57BL/6Jmice that were treated with SW033291 5 mg/kg IP daily for 3 doses in avehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W or that were treatedwith vehicle alone. On day 3 mice were sacrificed, marrow flushed andmixed at a 1:1 ratio with vehicle treated CD45.1 marrow. 2 million wholeBM cells were injected into the tail vein of lethally irradiated CD45.1mice and percent chimerism measured via flow cytometry at weeks 8, 12,16. As shown, at weeks 12 and 16 the percent blood chimerism of CD45.2marked cells was significantly increased in recipient mice whose CD45.2marked marrow was harvested from SW033291 treated donor mice, as opposedto vehicle control treated donor mice. In other words, marrow fromSW033291 treated mice demonstrated long term increased fitness incompetition with control marrow. In particular, at week 16 CD45.2harvested from SW033291 treated mice show a significant increase incontribution to B and T cell populations, suggesting marrow fromSW033291 treated mice promotes earlier reconstitution of lymphoidpopulations and earlier return to immune competence.

In an additional study, C57BL/6J mice are irradiated with 11Gy on day 0,followed by treatment with SW033291 5 mg/kg IP twice daily (bid) in avehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W, or with vehicle onlyfor 21 days. Mice treated with vehicle or with SW033291 all receive anallograft of marrow from a donor C57BL/6J mouse at a dose of either100,000 cells, 200,000 cells, 500,000 cells. 3 control and 3 SW033291mice are assessed under each condition. The experimental design isdepicted in FIG. 50.

Table 1 shows the number of surviving mice in each cohort over the first19 days of study. Under the conditions of the mouse colony during thisstudy, control mice receiving 100,000-500,000 cells are all dead betweendays 4-13 of study. In contrast, two SW033291 treated mice receiving500,000 cells remain alive on day 19 of the study and are presumed tohave full hematopoietic reconstitution. Thus treatment with the 15-PGDHinhibitor SW033291 promoted survival of mice receiving a bone marrowtransplant, an observation consistent with SW033291 enabling more rapidand complete hematopoietic reconstitution in the transplanted mice.Other 15-PGDH inhibitors with activity similar to SW033291 would bepredicted to have similar activity in supporting hematopoieticreconstitution. Treatment with SW033291 also enabled mice to besuccessfully transplanted with a smaller inoculum of donor bone marrowthan the 1,000,000 cells that are standardly needed. These observationssuggest SW033291, as well as other similar 15-PGDH inhibitors, is ableto support successful transplantation with smaller numbers of donor stemcells. Such activity would be of particular utility in settings, such astransplantation with umbilical cord stem cells, in which donor cellnumbers are limited. Improved survival of transplanted mice treated withSW033291 suggests efficacy of SW033291, and of similar 15-PGDHinhibitors, as replacements for, or in enabling decreased use of, othertreatments or growth factors commonly employed in support of patientsreceiving bone marrow, hematopoietic stem cell, and cord blood stem celltransplants. Improved survival of transplanted mice treated withSW033291 is consistent with SW033291, and by extension other similar15-PGDH inhibitors, having activity in reducing infections in thetransplanted mice, and/or in promoting recovery of mice intestines fromdamage by radiation, and/or in reducing pulmonary toxicity fromradiation.

TABLE 1 Mouse 13- 14- 15- 16- 17- 18- 19- 20- 21- 22- 23- 24- 25- 26- 1-survival Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar AprCell Day Day Day Day Day Day Day Day Day Day Day Day Day Day DayTreatment number 0 1 2 3 4 5 6 7 8 9 10 11 12 13 19 Control1×10{circumflex over ( )}5 3 3 3 2 0 Control 2×10{circumflex over ( )}53 3 3 3 3 2 1 0 Control 5×10{circumflex over ( )}5 3 3 3 3 3 3 2 1 1 1 11 1 0 SW033291 1×10{circumflex over ( )}5 3 3 3 3 2 1 0 SW0332912×10{circumflex over ( )}5 3 3 3 3 3 3 2 2 2 2 2 2 1 0 SW0332915×10{circumflex over ( )}5 3 3 3 3 3 3 3 3 2 2 2 2 2 2 . . . 2

Example 2

Analysis of Effect of SW033291 on Radiation Survival

This Example shows studies of the effect of SW033291 in mice receivingwhole body irradiation.

Table 2 shows the results of a study of 15 week old C57BL/6J female miceirradiated with 7Gy, 9Gy, or 11Gy, and receiving daily SW033291 5 mg/kgIP in a vehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W for 7 doses, orreceiving vehicle alone. The table shows the number of mice surviving onsequential days of the study. Under the conditions of the mouse colonyduring this experiment, mice receiving a lethal dose of 11Gy lived 48hours longer if treated with SW033291 than if receiving vehicle control,with control mice all dead on day 8; whereas SW033219 treated mice wereall dead on day 10.

TABLE 2 10/2 10/3 10/4 10/5 10/6 10/7 10/8 10/9 10/10 10/11 10/12 10/1310/23 Radiation Treatment Day Day Day Day Day Day Day Day Day Day DayDay Day Day Dose Arm 0 1 2 3 4 5 6 7 8 9 14 15 16 25 7 Gy Saline 3 3 3 33 3 3 3 3 3 3 3 Looks Healthy SW033291 3 3 3 3 3 3 3 3 3 3 3 3 LooksHealthy 9 Gy Saline 3 3 3 3 3 3 3 3 2 1 1 0 SW033291 3 3 3 3 3 3 3 3 2 21 0 11 Gy Saline 3 3 3 2 0 SW033291 3 3 3 3 3 2 0

Table 3 shows the number of mice surviving on sequential days of a studyof mice treated at 11Gy treated with either vehicle control or withSW033291 IP, in a vehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W, withSW033291 administered either at 5 mg/kg daily for 7 days, 5 mg/kg dailythroughout the study, or at 5 mg/kg twice daily for 7 days. Again micetreated with SW033291 on any of these dosing schedules live on average1-2 days longer than mice receiving vehicle control. The activity ofSW033291 in promoting resistance to toxic effects of radiation mayextend to SW033291 and other similar 15-PGDH inhibitors in promotingresistance to other similar toxic insults including but not limited toCytoxan, fludarabine, chemotherapy and immunosuppressive therapy.

TABLE 3 Friday Wed. Thurs. Friday Saturday Sunday Monday Treatment12-Oct 17-Oct 18-Oct 19-Oct 20-Oct 21-Oct 22-Oct Conditions Day 0 Day 5Day 6 Day 7 Day 8 Day 9 Day10 11 Saline (7 2 2 2 2 0 Gy days, 1 dosedaily) 11 5W033291 3 3 3 3 2 1 0 Gy (1 dose/daily) for 7 days 115W033291 3 3 3 3 3 2 0 Gy (1 dose/daily, continuous every day) 11 Saline(7 3 3 3 2 1 0 0 Gy days, 2 dose/daily 11 SW033291 (7 3 3 3 3 3 2 0 Gydays, 2 dose daily)

Example 3

Analysis of Effect of SW033291 on Survival of Mice Following Bone MarrowTransplant

FIG. 5 shows enhanced survival in mice receiving a bone marrowtransplant and also administered the 15-PGDH inhibitor SW033291. FIG. 5Ashows the design of the study. Mice were irradiated with a bone marrowablative dose of 11Gy on day 0, followed by administration of SW033291in 5 mg/kg twice daily by intraperitoneal injection in a vehicle of 10%Ethanol, 5% Cremophor EL, 85% D5W at a concentration of 125 μg/200 μl. Amatched control cohort received injections with vehicle only. On day onemice received an infusion of donor marrow at doses of 100,000 cells;200,000 cells; or 500,000 cells. FIG. 5B shows graphical survival curvesfor mice transplanted with 100,000 donor cells. FIG. 5C shows graphicalsurvival curves for mice transplanted with 200,000 donor cells. FIG. 5Dshows graphical survival curves for mice transplanted with 500,000 donorcells. Table 4 shows tabular survival data for all mice in the study onstudy day 30. Among mice receiving 100,000 donor cells, all micesuccumb, but SW033291 treated mice show an approximate doubling ofmedian survival. Among mice receiving 200,000 donor cells, all controlmice were dead by day 12. In contrast, all mice receiving 200,000 donorcells plus SW033291 survived at 30 days of observation and weresuccessfully engrafted. Among mice receiving 500,000 donor cells,control mice showed a 37.5% mortality; whereas, mice receiving SW033291again all survived.

TABLE 4 Dose Vehicle Survival SW033291 Survival 1 * 10⁵ 0/8 1/8 2 * 10⁵0/8 8/8 5 * 10⁵ 5/8 8/8

FIG. 6 shows a set of studies conducted on lethally irradiated mice thatreceived 500,000 donor marrow cells and were treated with eitherSW033291 at 5 mg/kg intraperitoneal dose twice daily or with vehiclecontrol, in a design otherwise identical to that used for the studies ofFIG. 5. FIG. 6A shows measurements on blood and bone marrow on day 5after transplant, with FIG. 6B showing that SW033291 treated mice havesignificantly higher total white count and FIG. 6C showing that SW033291treated mice have significantly higher total platelet count. The starsymbol denotes P<0.05.

FIG. 7A shows measurements on blood and bone marrow on day 8 aftertransplant. FIG. 7B shows that SW033291 treated mice have significantlyhigher platelet count than control, with drug treated mice having 77,000platelets compared to control mice having 39,500 platelets. The starsymbol denotes P<0.05.

FIG. 8A shows measurements on blood and bone marrow on day 12 aftertransplant. FIG. 8B shows that SW033291 treated mice have significantlyhigher neutrophil counts, with drug treated mice having 332 neutrophilscompared to control mice having 125 neutrophils. FIG. 8C shows that onday 12 after transplant, SW033291 treated mice have significantly higherhemoglobin count than controls, with drug treated mice having hemoglobinlevel of 11.58 and control mice having hemoglobin level of 8.3.Additionally, FIG. 8D shows SW033291 treated mice also havesignificantly greater total white counts than control mice. The starsymbol denotes P<0.05.

FIG. 9A shows measurements on blood and bone marrow on day 18 aftertransplant. SW033291 treated mice have significantly higher total whitecount (FIG. 9B), lymphocyte count (FIG. 9C), and neutrophil count (FIG.9D), with drug treated mice having 835 neutrophils and control micehaving 365 neutrophils (FIG. 9D). In comparison with counts on day 12,administration of SW033291 accelerates recovery of neutrophil counts bynearly 6 days (FIG. 8B versus FIG. 9D). FIG. 8E also shows that on day18 drug treated mice have significantly higher platelet counts thancontrol mice. Last, day 18, drug treated mice have nearly 4-foldincreased percentage (FIG. 8F) and total numbers (FIG. 9G) of SKL markedbone marrow stem cells than do control mice, with drug treated micehaving a mean of 4127 SKL marked bone marrow cells compared to controlmice having a mean of 967 SKL marked bone marrow cells. The star symboldenotes P<0.05.

FIG. 10(A) shows measurement of PGE2 (pg of PGE2/mg tissue protein) in 4different mouse tissues (colon, bone marrow, liver, lung) across timefollowing IP injection of SW033291 at 10 mg/kg. Blue bar representsbaseline at time 0, and red bars represent time course of PGE2concentration from 1-12 hours following SW033291 injection. FIG. 10(B)shows time course of PGE2 in control mice injected with vehicle only.

FIG. 11 shows a schema of an experiment in which mice are lethallyirradiated (IR) and 12 hours later receive a transplant (BMT) with CFSEdye labeled bone marrow cells (BM), and the numbers of transplantedcells that home and survive in the bone marrow of the recipient mice arethen determined by FACS at 16 hours post-transplant. In different armsof the experiment mice are treated with vehicle, with SW033291 (10 mg/kgIP), or with SW033291 (10 mg/kg IP) plus Indomethacin. Drugs areadministered following radiation, following the transplant, and again at8 hours after the transplant.

FIG. 12 shows a graph illustrating the percent of CFSE dye labeled cellsthat have homed to the bone marrow of mice treated as per the schemadescribed in FIG. 11. Treating mice with SW033291 concurrent with andfollowing the bone marrow transplant increases numbers of homed cells inthe recipient mouse bone marrow 3-fold. The figure further shows thatthe effect of SW033291 is near completely blocked by indomethacin(Indo+SW0), an inhibitor of COX enzymes the produce prostaglandins. Thisis consistent with the effect of SW033291 being mediated through theinhibition of 15-PGDH and through the resulting increase in tissueprostaglandins.

FIG. 13 shows a schema of an experiment in which mice are lethallyirradiated (IR) and 12 hours later receive a transplant (BMT) with CFSEdye labeled bone marrow cells (BM), and numbers of transplanted cellsthat home and survive in the bone marrow of the recipient mice are thendetermined by FACS at 16 hours post-transplant. In different arms of theexperiment mice are treated with vehicle, with SW033291 (10 mg/kg IP),with SW033291 (10 mg/kg IP) plus an antagonist of PGE2 receptor EP2(PF-04418948), or with SW033291 (10 mg/kg IP) plus an antagonist of PGE2receptor EP4 (L-161,982). Drugs are administered following radiation,following the transplant, and again at 8 hours after the transplant.

FIG. 14 shows a graph illustrating the percent of CFSE dye labeled cellsthat have homed to the bone marrow of mice treated as per the schemadescribed in FIG. 13. Treating mice with SW033291 concurrent with andfollowing the bone marrow transplant increases numbers of homed cells inthe recipient mouse bone marrow 2-fold. The figure further shows thatthe effect of SW033291 is near completely blocked by an antagonist tothe EP4 receptor (EP4+SW0), and is partially blocked by an antagonist ofthe EP2 receptor (EP2+SW0). This is consistent with the effect ofSW033291 being mediated through the inhibition of 15-PGDH and throughthe resulting increase of tissue prostaglandins, including PGE2.

In another experiment, mice were injected with SW033291 twice daily IPat 10 mg/kg for 5 doses. 2 hours following the last dose bone marrow isharvested and sorted into SKL marked cells that are hematopoietic stemcell enriched and into CD45 (−) cells that are bone marrow stroma cells.RNA was extracted and gene expression determined relative to levels inmice injected with vehicle control.

FIGS. 15(A-B) show graph illustrating induction of gene expression inbone marrow SKL cells and bone marrow stromal cells of SW033291 treatedmice. Mice administered SW033291 show a 3 fold induction in RNAexpression of CXCL12 and SCF in bone marrow SKL cells, and show agreater than 4-fold induction of CXCL12 and SCF in CD45(−) bone marrowstromal cells.

FIG. 16 illustrates a schema of an experiment in which immune deficientNSG mice are lethally irradiated (IR) and 12 hours later receive atransplant with CFSE dye labeled buffy coat cells from human umbilicalcord blood (UCB), and number of transplanted cells that home and survivein the bone marrow of the recipient mice are then determined by FACS at16 hours post-transplant. In different arms of the experiment mice aretreated with vehicle or with SW033291. Drugs are administered followingradiation, following the transplant, and again at 8 hours after thetransplant.

FIG. 17 illustrates a graph showing the percent of CFSE dye labeledhuman umbilical cord buffy coat cells that have homed to the bone marrowof mice treated as per the schema described in FIG. 18. Treating micewith SW033291 at the time of and following the transplant with buffycoat from human umbilical cord blood (UCB) increases numbers of homedhuman cells in the recipient mouse bone marrow nearly 2-fold.

Example 3

FIG. 18 shows the design of a study in which 4 groups of mice (4 miceper group) were administered either;

a) vehicle control,

b) 2.5 mg/kg of 15-PGDH inhibitor (+) SW209415 twice daily IP for 7doses,

c) recombinant human G-CSF 250 ug/kg subcutaneously once daily for 4doses

d) the combination of 2.5 mg/kg of 15-PGDH inhibitor (+) SW209415 twicedaily IP for 7 doses plus recombinant human G-CSF 250 ug/kgsubcutaneously once daily for 4 doses (with the daily dose of G-CSFbeing administered coincident with a dose of (+) SW209415) 3 hoursfollowing the last drug dose mice were sacrificed for analysis.

FIG. 19 shows results in each arm for measurement of total white cells,neutrophils, and lymphocytes.

Neutrophil counts approximately double following administration ofG-CSF, approximately doubled following administration of (+) SW209415,and increased 3 to 4-fold after administration of the combination ofG-CSF plus (+) SW209415.

FIG. 20 shows results in each arm for measurement of red cells,hematocrit, hemoglobin, and platelets.

FIG. 21 shows results in each arm for measurement in peripheral blood ofcirculating SKL (Sca-1+; C-kit+; Lin−) marked cells that are enrichedfor hematopoietic stem cells. The results show first that in this studyG-CSF increased the frequency of peripheral blood SKL cells (in Lin−cells) by 33%. (+) SW209415 when administered alone showed no effect onfrequency of peripheral blood stem cells. However, the combination ofG-CSF plus (+) SW209415 increased by 2 to 3-fold the frequency ofperipheral blood SKL cells (in Lin− cells). This result understates theabsolute magnitude of this effect, as the combination of G-SCF plus (+)SW209415 also increases the total white count, and hence the absolutenumber of the Lin− population. Thus, G-CSF increased the number ofperipheral blood SKL cells by approximately 50%. (+) SW209415 whenadministered alone showed no effect on number of peripheral blood stemcells. However, the combination of G-CSF plus (+) SW209415 increased thenumber of peripheral blood SKL cells by approximately 3.5-fold, showinga marked increased effect of the combination.

FIG. 22 shows results in each arm for measurement of SKL cells in micebone marrow. The show that bone marrow SKL marked cells are increased by(+) SW209415, further increased by G-CSF, and further increased by thecombination of (+) SW209415 plus G-CSF.

Example 4

In this Example, recipient mice lethally irradiated (11 Gy) followed byBMT (500 k total cells) 16 hours post IR were administered either:

a) Vehicle treated (2× daily).

b) 250 μg/kg GCSF (1× daily, subcutaneous administration, starting 24hours post BMT)

c) 2.5 mg/kg SW209415 (+) (2× daily, 18 days, starting post IR).

d) 250 μg/kg hGCSF (lx daily, 17 days)+2.5 mg/kg SW209415(+) (2× daily,18 days).

Mice were bled via submandibular vein at days 8, 12, and 18 posttransplant for CBC using the Hemavet 850 fs and sacrificed at day 18 toassess marrow cellularity and SKL frequency. Blood counts were tabulatedgraphically as illustrated in FIGS. 24-26 with error bars correspondingto standard error of the means and compared using 2-tailed t-tests.

SW209415 is Synergistic with GCSF in Promoting Hematopoietic Recoveryafter BMT

Granulocyte colony stimulating factor (G-CSF, clinical names Lenograstimand Filgrastim) is an FDA approved glycoprotein used bothpre-transplantation for stem cell priming purposes, and posttransplantation to enhance stem cell engraftment and minimize mortalityassociated with prolonged neutropenia. Its use has been shown toaccelerate granulocyte engraftment by 1-6 days following BMT compared tocontrol. G-CSF occurs naturally and is responsible for mounting theneutrophil activation in response to stress or infection by binding toits receptor G-CSFR. Several tissues can produce G-CSF upon stimulationby inflammatory mediators such as LPS, TNF-α, and IL-17, resulting inG-CSF release into the bloodstream which stimulates neutrophilproduction within, and mobilization from the bone marrow. In addition toneutrophil mobilization, G-CSF has been implication in mobilizing HSCsfrom marrow.

The use of SW209415 clinically in recipients of BMT would needvalidation in clinical models in comparison and in combination withGCSF. To determine SW209415's ability to promote hematopoietic recoveryfollowing BMT independently as well as synergistically with GCSF, weestablished the bone marrow transplant model described above in whichrecipient mice receiving 500 k total bone marrow cells were assessed forperipheral blood recovery at 3 time points post transplant. FIGS. 23-26show that compared to vehicle treatment, W209415, GCSF, and thecombination treatment recovered neutrophils significantly faster at Days8, 12, and 18 post BMT, but that the combination treatment demonstrateda synergistic effect. Neutrophil counts at Day 8-150 (Veh), 339(415),295(GCSF), 466-(4+G), Day 12-427(Veh) 839 (415), 982 (GCSF), 1390 (4+G),and Day 18-649 (Veh), 1370 (415), 1518 (GCSF), 1703 (4+G) alldemonstrated an enhanced effect when the agents were used incombination, showing the two compounds promote recovery via independentmechanisms. Additionally we showed the frequency of SKL cells (enrichedfor true HSCs) was higher in the combination treated group compared toeither therapeutic alone.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims. All patents, publications andreferences cited in the foregoing specification are herein incorporatedby reference in their entirety.

The following is claimed:
 1. A method of increasing neutrophils in asubject in need thereof, the method comprising administering to thesubject a 15-PGDH inhibitor, wherein the 15-PGDH inhibitor has thefollowing formula (V):

wherein n is 0-2 X⁶ is independently N or CR^(c) R¹, R⁶, R⁷, and R^(c)are each independently selected from the group consisting of hydrogen,substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl, C₂-C₂₄alkynyl, C₃-C₂₀ aryl, heteroaryl, heterocycloalkenyl containing from 5-6ring atoms, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃ alkyl)₃,hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄alkynyloxy, C₅-C₂₀ aryloxy, acyl, acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀aryloxycarbonyl, C₂-C₂₄ alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy,carboxylato, carbamoyl, C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl,thiocarbamoyl, carbamido, cyano, isocyano, cyanato, isocyanato,isothiocyanato, azido, formyl, thioformyl, amino, C₁-C₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido, C₆-C₂₀ arylamido, imino,alkylimino, arylimino, nitro, nitroso, sulfo, sulfonato, C₁-C₂₄alkylsulfanyl, arylsulfanyl, C₁-C₂₄ alkylsulfinyl, C₅-C₂₀ arylsulfinyl,C₁-C₂₄ alkylsulfonyl, C₅-C₂₀ arylsulfonyl, sulfonamide, phosphono,phosphonato, phosphinato, phospho, phosphino, polyalkylethers,phosphates, and phosphate esters, and wherein R⁶ and R⁷ may be linked toform a cyclic or polycyclic ring, wherein the ring is a substituted orunsubstituted aryl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted cycloalkyl, and a substituted orunsubstituted heterocyclyl; U¹ is N, C—R², or C—NR³R⁴, wherein R² isselected from the group consisting of a H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X,CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X, (wherein X═H, F, Cl, Br, or I), CN,(C═O)—R′, (C═O)N(R′)₂, O(CO)R′, COOR′ (wherein R′ is H or a lower alkylgroup), and wherein R¹ and R² may be linked to form a cyclic orpolycyclic ring, wherein R³ and R⁴ are the same or different and areeach selected from the group consisting of H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, CH₂—CH₂—CH₂X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(R′)₂, COOR′(wherein R′ is H or a lower alkyl group), and R³ or R⁴ may be absent;and pharmaceutically acceptable salts thereof.
 2. The method of claim 1,further comprising administering in combination with the 15-PGDHinhibitor at least one hematopoietic cytokine or cell mobilizationagent.
 3. The method of claim 1, wherein the hematopoietic cytokine orcell mobilization agent comprises at least one of G-CSF or Plerixafor.4. The method of claim 1, wherein the subject has or is at risk of atleast one of neutropenia, thrombocytopenia, anemia, or cytopenia.
 5. Themethod of claim 4, wherein the subject has received a hematopoietic stemcell transplant, bone marrow transplant, chemotherapy, amyelosuppressive therapy, radiation therapy, and/or viral therapy,and/or has a bone marrow disease, cancer, viral infection, aplasticanemia, myelodysplasia, and/or myelofibrosis.
 6. The method of claim 1,wherein the 15-PGDH inhibitor has the following formula (VI):

wherein n=0-2; X⁶ is N or CR^(c); R¹ is selected from the groupconsisting of branched or linear alkyl, —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5); R⁵ is selected from the group consisting of H, Cl, F, NH₂, andN(R⁷⁶)₂; R⁶ and R⁷ can each independently be one of the following:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, R⁷⁴, R⁷⁶, and R^(c)are the same or different and are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl,acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀ aryloxycarbonyl, C₂-C₂₄alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy, carboxylato, carbamoyl,C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,thioformyl, amino, C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄alkylamido, C₆-C₂₀ arylamido, imino, alkylimino, arylimino, nitro,nitroso, sulfo, sulfonato, C₁-C₂₄ alkylsulfanyl, arylsulfanyl, C₁-C₂₄alkylsulfinyl, C₅-C₂₀ arylsulfinyl, C₁-C₂₄ alkylsulfonyl, C₅-C₂₀arylsulfonyl, sulfonamide, phosphono, phosphonato, phosphinato, phospho,phosphino, polyalkylethers, phosphates, phosphate esters, andpharmaceutically acceptable salts thereof.
 7. The method of claim 1,wherein the 15-PGDH inhibitor has the following formula followingformula:

or pharmaceutically acceptable salts thereof.
 8. A method increasingnumbers of and/or of mobilizing peripheral blood hematopoietic stemcells in a subject in need thereof, the method comprising administeringto the subject a 15-PGDH inhibitor, wherein the 15-PGDH inhibitor hasthe following formula (V):

wherein n is 0-2 X⁶ is independently is N or CR^(c) R¹, R⁶, R⁷, andR^(c) are each independently selected from the group consisting ofhydrogen, substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl, heterocycloalkenyl containingfrom 5-6 ring atoms, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄alkynyloxy, C₅-C₂₀ aryloxy, acyl, acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀aryloxycarbonyl, C₂-C₂₄ alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy,carboxylato, carbamoyl, C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl,thiocarbamoyl, carbamido, cyano, isocyano, cyanato, isocyanato,isothiocyanato, azido, formyl, thioformyl, amino, C₁-C₂₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido, C₆-C₂₀ arylamido, imino,alkylimino, arylimino, nitro, nitroso, sulfo, sulfonato, C₁-C₂₄alkylsulfanyl, arylsulfanyl, C₁-C₂₄ alkylsulfinyl, C₅-C₂₀ arylsulfinyl,C₁-C₂₄ alkylsulfonyl, C₅-C₂₀ arylsulfonyl, sulfonamide, phosphono,phosphonato, phosphinato, phospho, phosphino, polyalkylethers,phosphates, and phosphate esters, and wherein R⁶ and R⁷ may be linked toform a cyclic or polycyclic ring, wherein the ring is a substituted orunsubstituted aryl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted cycloalkyl, and a substituted orunsubstituted heterocyclyl; U¹ is N, C—R², or C—NR³R⁴, wherein R² isselected from the group consisting of a H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X,CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X, (wherein X═H, F, Cl, Br, or I), CN,(C═O)—R′, (C═O)N(R′)₂, O(CO)R′, COOR′ (wherein R′ is H or a lower alkylgroup), and wherein R¹ and R² may be linked to form a cyclic orpolycyclic ring, wherein R³ and R⁴ are the same or different and areeach selected from the group consisting of H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, CH₂—CH₂—CH₂X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(F)₂, COOR′ (whereinR′ is H or a lower alkyl group), and R³ or R⁴ may be absent; andpharmaceutically acceptable salts thereof.
 9. The method of claim 8,wherein the number of hematopoietic stem cells is increased in blood orbone marrow.
 10. The method of claim 8, further comprising administeringin combination with the 15-PGDH inhibitor at least one hematopoieticcytokine or cell mobilization agent.
 11. The method of claim 10, whereinthe hematopoietic cytokine or cell mobilization agent comprises at leastone of G-CSF or Plerixafor.
 12. The method of claim 8, wherein thesubject has or is at risk of at least one of neutropenia,thrombocytopenia, anemia, or cytopenia.
 13. The method of claim 12,wherein the subject has received a hematopoietic stem cell transplant,bone marrow transplant, chemotherapy, a myelosuppressive therapy,radiation therapy, viral therapy, and/or has a bone marrow disease,cancer, viral infection, aplastic anemia, myelodysplasia, and/ormyelofibrosis.
 14. The method of claim 8, wherein the 15-PGDH inhibitorhas the following formula (VI):

wherein n=0-2; X⁶ is N or CR^(c); R¹ is selected from the groupconsisting of branched or linear alkyl —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5); R⁵ is selected from the group consisting of H, Cl, F, NH₂, andN(R⁷⁶)₂; R⁶ and R⁷ can each independently be one of the following:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³,R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, R⁷⁴, R⁷⁶, and R^(c)are the same or different and are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl,acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀ aryloxycarbonyl, C₂-C₂₄alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy, carboxylato, carbamoyl,C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,thioformyl, amino, C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄alkylamido, C₆-C₂₀ arylamido, imino, alkylimino, arylimino, nitro,nitroso, sulfo, sulfonato, C₁-C₂₄ alkylsulfanyl, arylsulfanyl, C₁-C₂₄alkylsulfinyl, C₅-C₂₀ arylsulfinyl, C₁-C₂₄ alkylsulfonyl, C₅-C₂₀arylsulfonyl, sulfonamide, phosphono, phosphonato, phosphinato, phospho,phosphino, polyalkylethers, phosphates, phosphate esters, andpharmaceutically acceptable salts thereof.
 15. The method of claim 8,wherein the 15-PGDH inhibitor has the following formula followingformula:

or pharmaceutically acceptable salts thereof.
 16. A method of treatingat least one of neutropenia, thrombocytopenia, anemia, or cytopenia in asubject in need thereof, the method comprising: administering to thesubject a 15-PGDH inhibitor, wherein the 15-PGDH inhibitor has thefollowing formula (V):

wherein n is 0-2 X⁶ is independently is N or CR^(c) R¹, R⁶, R⁷, andR^(c) are each independently selected from the group consisting ofhydrogen, substituted or unsubstituted C₁-C₂₄ alkyl, C₂-C₂₄ alkenyl,C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl, heterocycloalkenyl containingfrom 5-6 ring atoms, C₆-C₂₄ alkaryl, C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄alkynyloxy, C₅-C₂₀ aryloxy, acyl, acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀aryloxycarbonyl, C₂-C₂₄ alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy,carboxylato, carbamoyl, C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl,thiocarbamoyl, carbamido, cyano, isocyano, cyanato, isocyanato,isothiocyanato, azido, formyl, thioformyl, amino, C₁-C₄ alkyl amino,C₅-C₂₀ aryl amino, C₂-C₂₄ alkylamido, C₆-C₂₀ arylamido, imino,alkylimino, arylimino, nitro, nitroso, sulfo, sulfonato, C₁-C₂₄alkylsulfanyl, arylsulfanyl, C₁-C₂₄ alkylsulfinyl, C₅-C₂₀ arylsulfinyl,C₁-C₂₄ alkylsulfonyl, C₅-C₂₀ arylsulfonyl, sulfonamide, phosphono,phosphonato, phosphinato, phospho, phosphino, polyalkylethers, andphosphates, phosphate esters, and wherein R⁶ and R⁷ may be linked toform a cyclic or polycyclic ring, wherein the ring is a substituted orunsubstituted aryl, a substituted or unsubstituted heteroaryl, asubstituted or unsubstituted cycloalkyl, and a substituted orunsubstituted heterocyclyl; U¹ is N, C—R², or C—NR³R⁴, wherein R² isselected from the group consisting of a H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, O—CH₂—CH₂X,CH₂—CH₂—CH₂X, O—CH₂—CH₂X, X, (wherein X═H, F, Cl, Br, or I), CN,(C═O)—R′, (C═O)N(R′)₂, O(CO)R′, COOR′ (wherein R′ is H or a lower alkylgroup), and wherein R¹ and R² may be linked to form a cyclic orpolycyclic ring, wherein R³ and R⁴ are the same or different and areeach selected from the group consisting of H, a lower alkyl group, O,(CH₂)_(n1)OR′ (wherein n1=1, 2, or 3), CF₃, CH₂—CH₂X, CH₂—CH₂—CH₂X,(wherein X═H, F, Cl, Br, or I), CN, (C═O)—R′, (C═O)N(F)₂, COOR′ (whereinR′ is H or a lower alkyl group), and R³ or R⁴ may be absent; andpharmaceutically acceptable salts thereof.
 17. The method of claim 16,further comprising administering in combination with the 15-PGDHinhibitor at least one hematopoietic cytokine or cell mobilizationagent.
 18. The method of claim 17, wherein the hematopoietic cytokine orcell mobilization agent comprises at least one of G-CSF or Plerixafor.19. The method of claim 16, wherein the subject has received ahematopoietic stem cell transplant, bone marrow transplant,chemotherapy, a myelosuppressive therapy, radiation therapy, and/orviral therapy, and/or has a bone marrow disease, cancer, viralinfection, aplastic anemia, myelodysplasia, and/or myelofibrosis. 20.The method of claim 16, wherein the 15-PGDH inhibitor has the followingformula (VI):

wherein n=0-2; X⁶ is N or CR^(c); R¹ is selected from the groupconsisting of branched or linear alkyl, —(CH₂)n₁CH₃ (n₁=0-7),

wherein n₂=0-6 and X is any of the following: CF_(y)H_(z) (y+z=3),CCl_(y)H_(z) (y+z=3), OH, OAc, OMe, R⁷¹, OR⁷², CN, N(R⁷³)₂,

(n₃=0-5, m=1-5), and

(n₄=0-5); R⁵ is selected from the group consisting of H, Cl, F, NH₂, andN(R⁷⁶)₂; R⁶ and R⁷ can each independently be one of the following:

each R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹,R²², R²³, R²⁴, R²⁵, R²⁶, R²⁷, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, R³⁴, R³⁵,R³⁶, R³⁷, R³⁸, R³⁹, R⁴⁰, R⁴¹, R⁴², R⁴³, R⁴⁴, R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹,R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁶, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², R⁶³R⁶⁴, R⁶⁵, R⁶⁶, R⁶⁷, R⁶⁸, R⁶⁹, R⁷⁰, R⁷¹, R⁷², R⁷³, R⁷⁴, R⁷⁶, and R^(c)are the same or different and are independently selected from the groupconsisting of hydrogen, substituted or unsubstituted C₁-C₂₄ alkyl,C₂-C₂₄ alkenyl, C₂-C₂₄ alkynyl, C₃-C₂₀ aryl, heteroaryl,heterocycloalkenyl containing from 5-6 ring atoms, C₆-C₂₄ alkaryl,C₆-C₂₄ aralkyl, halo, —Si(C₁-C₃ alkyl)₃, hydroxyl, sulfhydryl, C₁-C₂₄alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy, acyl,acyloxy, C₂-C₂₄ alkoxycarbonyl, C₆-C₂₀ aryloxycarbonyl, C₂-C₂₄alkylcarbonato, C₆-C₂₀ arylcarbonato, carboxy, carboxylato, carbamoyl,C₁-C₂₄ alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,thioformyl, amino, C₁-C₂₄ alkyl amino, C₅-C₂₀ aryl amino, C₂-C₂₄alkylamido, C₆-C₂₀ arylamido, imino, alkylimino, arylimino, nitro,nitroso, sulfo, sulfonato, C₁-C₂₄ alkylsulfanyl, arylsulfanyl, C₁-C₂₄alkylsulfinyl, C₅-C₂₀ arylsulfinyl, C₁-C₂₄ alkylsulfonyl, C₅-C₂₀arylsulfonyl, sulfonamide, phosphono, phosphonato, phosphinato, phospho,phosphino, polyalkylethers, phosphates, phosphate esters, andpharmaceutically acceptable salts thereof.
 21. The method of claim 16,wherein the 15-PGDH inhibitor has the following formula followingformula:

or pharmaceutically acceptable salts thereof.